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Xuri™CellExpansionSystemW25 - GE Healthcare Life Sciences

1. OEO E F E ee ne Note As a default the tray will prepare for tilt at the end of the run This setting can be changed in System Settings Rocker Prepare for tilt at END 3 Grab the textured grip area on each side of the tray and in one movement pull the tray upwards and against you to position the tray into tilt position 4 Connect tubing from the Cellbag bioreactor to the harvest vessel 5 Empty the Cellbag bioreactor using a pump or gravity flow 6 Connect tubing from the Cellbag bioreactor to the harvest vessel Procedures after cultivation Follow the instructions below when the cultivation is finished Step Action 1 Clamp off the inlet and outlet gas filters of the Cellbag bioreactor 2 Disconnect the tubing from the inlet gas filter on the Cellbag bioreactor 3 Disconnect any other tubing and cables still connected to the Cellbag bioreactor 4 Release and remove the empty Cellbag bioreactor from the tray by pressing down the bag clamp opener 198 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 5 Perform cultivation 5 5 3 End cultivation Step Action 5 Dispose the Cellbag bioreactor according to GE Healthcare recommendations and local regulations 6 Turn off all gas supplies Shut down the system Follow the instructions below to shut down the system Step Action 1 Disconnect from system in UNICORN 2 Press the power button on the rocker front panel Th
2. Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 75 3 The UNICORN software 3 3 Methods in UNICORN 3 3 6 Scouting 3 3 6 Scouting Introduction Scouting is used to repeat a series of method runs automatically using different settings or with predetermined changes in the values for one or more Variables A Scouting scheme is defined as part of the method This chapter gives an overview of scouting and the scouting workflow and describes how to set up and edit a Scouting scheme Scouting is ideal for relatively simple variable combinations Set up a scouting scheme 76 The table below describes how to set up a Scouting scheme where a variable is varied In this example the rocking speed is varied Note The Start protocol will only be displayed before the first run in the Scouting experiment Step Action 1 Create a method and decide appropriate run parameters to be varied in the experiment The run parameters to be varied should be defined as Variables in your method See Section 3 3 2 Method creation on page 53 for information about how to create methods See Edit variables on page 68 for information about how to define new variables Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 6 Scouting Step Action 2 In the Method Editor e Click the Scouting icon in the toolbar Mo or e Select Tools Scouting Start Protocol
3. 262 xuri Cell Expansion System W25 User Manual 29 0646 22 AA PUMP 25 1A total time of ac tual shot 8 Reference information 8 8 Control settings 8 8 6 Pump control Description When the pump Is running for limited time this data represents the total time the pump will run View from RD CD PUMP 25 1B actual flow The estimated flow of the pump PUMP 25 1B actual speed The pump rpm measured by the pump itself PUMP 25 1B accumulated volume The estimated total volume since last reset PUMP 25 1B time left of actu al shot This is counter showing how much time is left until the pump stops when the pump is running for a limited time This is the case when the pump is started manually for a limited time during calibration or when used as acid base pump by pH control PUMP 25 1B total time of ac tual shot When the pump is running for limited time this data represents the total time the pump will run PUMP 25 2A actual flow The estimated flow of the pump PUMP 25 2A actual speed PUMP 25 2A accumulated volume The pump rpm measured by the pump itself The estimated total volume since last reset PUMP 25 2A time left of actu al shot This is counter showing how much time is left until the pump stops when the pump is running for a limited time This is the case when the pump is started manually for a limited time during calibration or when used as acid base pump b
4. Verification of CO Follow the instructions below to verify that CO is functioning correctly Step Action 1 Z Switch on the client computer and Switch on the rocker and wait for the system to initialize Start UNICORN and connect to system Make sure that the desired gas supplies are connected to the system Refer to Section 5 1 5 Connect gas to system on page 166 In the Process Picture click the Compressed Air button to select compressed air or the N2 button to select N gt Start the gas flow by clicking the right hand side of the Gas flow button xuri Cell Expansion System W25 User Manual 29 0646 22 AA 147 4 System control description 4 6 System verification Verification of O 148 Step Action 7 Enter a gas flow setpoint for example 0 2 I min in Settings Gas control Gas flow Setpoint Click OK 8 Enter a CO setpoint for example 5 in Settings Gas control CO2 Setpoint Click OK 9 Wait for a few minutes and verify that the Process Picture shows a correct measurement If the gas flow appears not to function correctly refer to Section 7 3 Xuri Cell Expansion W25 CBCU on page 212 Follow the instructions below to verify that O is functioning correctly Step Action 1 Switch on the client computer 2 Switch on the rocker and wait for the system to initialize 3 Start UNICORN and connect to system 4 Make sure that the desired gas supplies are connected to the system Refer
5. Result The Scouting dialog opens with the Scouting Variables dialog dis played on top Scouting Scouting parameters Scouting Variables Run Included C Duration C Gasflow setpoint C Heating setpoint C pH reading cycle time C Rocking Angle C Rocking Speed C Show details C Show unused variables Note When editing a scouting scheme only the Scouting dialog is displayed Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 77 3 The UNICORN software 3 3 Methods in UNICORN 5 5 6 SCOUTING Step Action 3 e Inthe Scouting Variables dialog select the appropriate variables to be varied by checking the appropriate boxes Check the Show details box if you want to display variables defined as detailed variables in your method These are rarely used as scouting variables Check the Show unused variables box if you want to display variables currently not used in the method Result The Scouting dialog is updated with the selected variables and their default values Scouting Scouting parameters Method Settings METHOD SETTINGS Run Included Rocking Speed RPM 1 20 0 It is possible to insert runs one by one see step 4 or insert series of runs see step 5 78 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 6 Scouting Step Action 4 To insert runs one by one e Inthe Scouting dialog select a row in th
6. New Method Ctri hy H Mew Method Queue A Open Ctrl o Close Result The New Method dialog opens Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 55 3 The UNICORN software 3 3 Methods in UNICORN 3 3 2 Method creation Step Action New Method e ystem 2 Create a new method by using the O Predefined Method VE 25 Template Empty Method Method Description Empty method to be used for creating text edited Methods for the Ready oProcess WAVE 25 systems In the New Method dialog e select a System e select the Empty Method radio button e click OK Result An empty method that consists of the mandatory Method Settings phase is created 56 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 3 Work with methods 3 3 3 Work with methods Open method The table below describes how to open an existing method in the database Step Action 1 In the Method Editor e Click the Open Method Navigator icon in the Toolbar pay or e select File Open Toolbar Method Navigator h Phase Library Result The Method Navigator is displayed Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 57 3 The UNICORN software 3 3 Methods in UNICORN 3 3 3 Work with methods Step Action 2 Select the method to be opened in the Folder name column Method Navigator Fo
7. Save the report in PDF format The generated report can be saved as a PDF file as described below Step Action 1 Perform steps 1 to 2 in the Generate and print the report on page 98 instruc tion above Click the Preview button to view the report in the Customize Report window Select File Save As PDF Result The Save As dialog opens e Browse for a folder and File name for the report e Click Save Result The report is created as a PDF file and saved in the location specified in the dialog xuri Cell Expansion System W25 User Manual 29 0646 22 AA 99 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format 3 4 5 Create a new report format Introduction This section describes how to create a new customized report format You can choose from a variety of objects to include in a report including charts methods documentation free text and more You can also place align and size the objects as you please Note Click the Preview Edit mode button to toggle between the Preview mode which is for view only and the Edit mode where you can edit the report items The editing actions in this section are only available in the Edit mode Open the Customize Report window The table below describes how to open the Customize Report in Edit mode to create a new report format Step Action 1 Open a result in the Evaluation module 2 e Select File Report or e Click the Report i
8. Description The pH bag sensor port is located on the underside of the bag Holds a sensor for online pH control of the cell culture Outlet gas filter with pressure control valve The filter prevents airborne particles of 0 2 micron or larger to enter the bag The pressure control valve maintains a constant overpressure inside the Cellbag bioreactor Inlet gas filter The filter removes airborne particles of 0 2 micron or larger from the gas flow before it enters the Cellbag bioreactor Addition port Addition port equipped with a Luer quick connector DO bag sensor port The DO bag sensor is located on the under side of the bag Holds a sensor for online dissolved oxygen control of the cell culture Cellbag rod The Cellbag rod fixes the Cellbag bioreactor to the tray Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 55 2 System description 2 5 Cellbag bioreactor Component Description CLAVE sam Sampling port equipped with a self sealing pling port luer fitting Addition port Addition port equipped with a Luer quick connector Addition harvest Addition harvest port equipped with a MPC port quick connector pH and DO sensors 34 The Cellbag bioreactor may be equipped with optical sensors for monitoring of pH and dissolved oxygen DO The sensors are light sensitive and should therefore be protected from excessive light The sensors are loca
9. F Fast fill Function verification 149 Feed pump Auto calibration 136 Foaming Reduction 143 G Gas flow Function verification 146 Operating conditions 142 Gas flow control settings 254 Gas mix Connect 167 Description 166 H Harvest waste pump Auto calibration 136 Heating control settings 252 Help Text instructions 64 Help utility 38 Instrument Components 230 Instrument configura tion 230 L Lid Size 20 Log on UNICORN 170 M Manual instructions 47 Media addition mode Principles 134 Media control Media addition mode 134 Perfusion mode 135 Media control settings 284 Method Editor Saving a method 72 Saving a phase 74 Method Editor module description 50 icons 51 Method notes Add to method 62 Edit 62 Method queues 82 Create 82 Mulitple systems 83 Methods Add notes 62 Create empty method 55 Create new method 53 Edit notes 62 Open a method 57 Saving 72 Saving a phase 74 Start protocol setup 61 Syntax errors in 230 unable to create predefined method 231 unable to export mehod to network drive 231 N Notes Method notes 62 Notes and tips 9 O O2 Function verification 148 O mix control settings 258 Operating conditions 141 Output data CO mix control 257 DO control O2 mode 281 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA DO control transition de lay 283 DO measurement 278 Gas flow control 255 Heatin
10. Spstern Fi Francine 51872012 1 24 17 Default a Gerald 51872012 1 25 5 Default Greta 51872012 1 235 Default Helen 1872012 1 242 Default E Hame Rubisco M onoGl Tris 1 System SystemC 2 e Select the folder in which to save the method e Entera method Name e Select for which System to save the method 4 Click Save Result The method is saved in the database Note For some systems an error message will appear if you are trying to save the method for e asystem using another instrument configuration and or another compo nent configuration than the method originally was created for and e the settings in the method depend on the component configuration It will still be possible to save the method but the phases in the method will be marked with an error symbol In order to be able to subsequently run the method either the method must be text edited or the component configuration of the system changed in the Administration module xuri Cell Expansion System W25 User Manual 29 0646 22 AA 73 3 The UNICORN software 3 3 Methods in UNICORN 3 3 5 Save a method Save a phase The instruction below describes how to save a phase to the Phase Library Step Action 1 Select the phase to be saved in the method outline Note A Method Settings phase cannot be saved as a separate phase with a new name If properties for the Method Settings phase are cha
11. The table below describes a problem and its solution Problem description Solution The export function is available when an archived result is selected Do not export an archived However it is not possible to export an archived result Instead the result result that was selected before the archived result will be exported Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 235 8 Reference information 8 Reference information In this chapter This chapter lists technical specifications and control settings for Xuri Cell Expansion System W25 The chapter also includes a list of wetted materials and a chemical resis tance guide This chapter contains the following sections Section See page 8 1 System specifications 237 8 2 Component specifications 238 8 3 Client computer specifications 242 8 4 Xuri Cell Expansion W25 CBCU semi wetted materials 244 8 5 Chemical resistance 245 8 6 Tubing and connectors 246 8 7 Auto calibration 247 8 8 Control settings 248 236 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 1 System specifications 8 1 System specifications The table below lists the system specifications of Xuri Cell Expansion System W25 Parameter System configuration Benchtop system external computer Control system Rocker embedded PC operating system UNICORN 6 3 2 or higher version Windows embedded standard 7 Connection between PC and inst
12. 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 Cautions CAUTION CAUTION indicates a hazardous situation which if not avoided could result in minor or moderate injury It is important not to pro ceed until all stated conditions are met and clearly understood Notices gt NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment 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 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 9 1 Introduction 1 2 Important concepts 1 2 Important concepts The concepts and abbreviations used in this manual are explained in the table below Concept Explanation abbreviation Cellbag bioreactor The disposable container in which the cells are cultured DO Dissolved oxygen DO sensor Optical sensor for measurement of dissolved oxygen At tached on DO configured Cellbag bioreactors pH sensor Optical sensor for pH measurement Attached on pH config ured Cellbag bioreactors CBCU Xuri Cell Expansion System W25 Cellbag Control Unit Refers to instrume
13. Zo 5 5 0 10 0 5 0 10 0 12 5 250 For the perfusion to work satisfactory it is important to ensure that the harvest waste filter and harvest waste out port is completely covered by liquid through the whole rocking cycle If the harvest waste out port is not always covered by liquid unnecessary warnings for harvest waste filter clogging might be given If auto calibration is used it will not calibrate the harvest waste pump correctly Emergency stop of pumps To avoid overfilling or draining the Cellbag due to for example lack of feed media har vest waste filter clogging or tubing error the feed or harvest waste pump will stop when the weight measurement deviates more than 10 from the weight setpoint The stopped pump will start again when the setpoint is reached 138 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 4 Media control 4 4 3 Deviation alarm 4 4 3 Deviation alarm By default a deviation alarm is activated It will send a message when the weight reading has been outside the specified toleration limits of the weight setpoint for a certain time The deviation alarm can be deactivated using the manual instruction Media control general During media addition the deviation alarm is acting on the momentary weight setpoint During perfusion the deviation alarm is active only in normal perfusion when both pumps are running Xuri Cell E
14. 0 00 Heating 37 0 Heating setpoint C On 0 5 C 1 0 C 60 sec 0 00 Gas flow 0 30 Gasflow setpoint l min On 0 05 l min 0 05 l min 60 sec CO2 mix 5 0 C02 setpoint On 1 0 1 0 60 sec 0 00 Set pH reading cycle time 120 pH reading cycle time sec 0 00 Start rocking 200 00 Duration End_Block Mi 0 00 Phase User Defined Instruction Bo Breakpoint Instructions 7 e a arameters for mix 0 00 hour oe A CO2 mix setpoint 0 0 15 0 amp Heating a ne a Var Gas flow 9 0 5 4 m mas Deviation alarm LU Mix Start CO2 mix Va jJ Off On _ Stop CO2 mix Allowed deviation up 1 0 5 0 02 mix la e pH sensor Var 1 0 7 pH control Allowed deviation down 1 0 5 0 H pH control advanced la ie DO sensor Var 1 0 2 DO control Deviation alarm delay 0 3600 DO control advanced a Media control Vat J 60 sec 5 Pump control manual y Result The New Variable dialog opens Mew Variable Variable name Po isible in details only Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 5 4 Text instructions Step Action 3 e Type a name for the variable e Select the Visible in details only checkbox if you want to set the variable as a detailed variable Detailed variables become visible in the Variable List if
15. Check that you have connected the inlet gas sup ply to the inlet filter does not have the pressure control valve Missing pressure con trol valve Check that the pressure control valve is present on the outlet filter The Cellbag bioreactor is underinflated and an alarm message in the pop up dialog indicates blockage of gas flow 228 Clogged inlet filter Check that the gas inlet flow path is unobstructed Tip A small amount of condensate in the inlet filter is normal However if excessive condensate is formed e Decrease the gas flow e Insulate the tubing to the pressure control valve Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 7 Troubleshooting 7 6 Software problems 7 6 Software problems In this section This section describes troubleshooting and corrective actions for UNICORN Refer to UNICORN Administration and Technical manual for more troubleshooting This section contains the following subsections Section See page 7 6 1 Troubleshooting Method editor 230 7 6 2 UNICORN System Control 252 7 6 3 Troubleshooting Evaluation 235 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 229 7 Troubleshooting 7 6 Software problems 7 6 1 Troubleshooting Method editor 7 6 1 Troubleshooting Method editor There are red instructions in a method Red instructions instructions with a red dot in a method are syntax errors and may have several causes A phase conta
16. Incorrect rocking angle Rocker is mechanically restricted from moving Localize and remove the restricting object Rocking speed varies DO control using speed is enabled If the DO deviates from the setpoint the rocking speed changes This is normal when DO control is enabled If not required disable DO control from the process pic ture 208 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Error symptom Power button is blink ing red Possible cause Rocker does not have any network connec tion to the UNICORN database System units such as CBCU or pump defined in the system setup in UNICORN are not con nected to the rocker 7 Troubleshooting 7 2 Xuri Cell Expansion System W25 rocker Corrective action Verify the Ethernet connection between the rocker and the client computer Disable the missing components in UNICORN See Section 5 3 Edit system properties on page 174 Connect the missing components to the rocker See Chapter 4 Installation in the Operating Instructions An internal error has occured in the embed ded computer of the rocker Read any warning message carefully and follow the instructions If problem persists contact GE Healthcare service personnel System does not shut down when the power button is pressed The embedded comput er of the rocker fails to shut down Wait one minute If the system still does not shut down keep p
17. Manual acid flow The acid flow used if Flow mode is set to Manual pH control advanced acid base Manual base flow The base flow used if Flow mode is set to Manual pH control advanced acid base Acid dead band The pH control output when using acid or base is a percentage of the cycle time telling how much of the cycle time the acid or base pump shall run The acid dead band defines the percentage interval in which the acid pump will not run l e if the acid dead band is 2 0 the output from the pH control needs to be above 2 0 for the acid pump to start pH control advanced acid base Base dead band The pH control output when using acid or base is a percentage of the cycle time telling how much of the cycle time the acid or base pump shall run The base dead band defines the percent age interval in which the base pump will not run l e if the base dead band is 2 0 the output from the pH control needs to be above 2 0 for the base pump to start pH control advanced acid base 1 This run data is not specifically connected to the acid or base role Instead this run data is connected to a specific pump head Look at the run data from the pump head of interest Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 213 8 Reference information 8 8 Control settings 8 8 8 pH control Note In the table the following abbreviations are used PP Process P
18. Start har mode will run as long as given by Har uous when vest waste vest pump duration If Continu started from pump ous is selected the pump will PP run until the harvest waste pump is stopped Harvest pump The time the harvest waste Start har duration pump will run if Harvest pump vest waste mode is set to Limited Else this pump parameter has no effect Harvestpump The rom at which the har Settings Cell Start har speed vest waste pump will run bag pumps vest waste pump 1 Select the pump head with the desired role acid base feed or harvest waste 2 Not shown in process picture when pump is used by pH or Media control Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Pump control output data Description View from od ey 1 D EEE C D PUMP 25 1A actual flow The estimated flow of the pump Y Y PUMP 25 1A actual speed The pump rpm measured by the pump itself yl jy PUMP 25 1A accumulated The estimated total volume since last reset Y Y y volume PUMP 25 1A time leftofactu This is counter showing how much time is left until Y al shot the pump stops when the pump is running for a limited time This is the case when the pump is started manually for a limited time during calibration or when used as acid base pump by pH control
19. tion alarm Select On to activate the devia tion alarm and Off to deactivate it Media control general Allowed devia tion up A parameter that defines how much above the setpoint the weight reading is allowed to be without triggering the deviation alarm Settings Me dia control Media control general Allowed devia tion down A parameter that defines how much below the setpoint the weight reading is allowed to be without triggering the deviation alarm Settings Me dia control Media control general Deviation alarm delay A parameter that defines for how long the weight reading can be outside the allowed deviation limits before the deviation alarm is triggered Media control general 284 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 11 Media control Setting Description Set from View from Flow rate The flow of the feed pump when media control is on In perfusion the harvest waste pump will al so start at this flow rate and then do minor flow adjustments to keep the weight set point Settings Me dia control Media control general PP D Auto calibra tion Select On to activate the auto calibration and Off to deactivate it Media control mode also needs to be set to Perfusion since no auto calibration will take place in Media addition When auto calibration is
20. If more than one pump unit is connected the units are distinguished by their CAN IDs The CAN ID is set by turning a switch on the pump rear panel see illustration above Four possible CAN ID positions are available on the switch marked 1 2 3 and 4 respec tively The CAN ID should be set to position 1 for the first pump and to position 2 for the second pump Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 31 2 System description 2 5 Cellbag bioreactor 2 5 Cellbag bioreactor Introduction Cell cultivation is performed inside the Cellbag bioreactor The Cellbag bioreactor is de livered gamma irradiated and ready for use It is intended for single use only and must be discarded after use Cellbag bioreactor options The Cellbag bioreactors are available in different configurations of varying sizes and equipped with various ports Cellbag bioreactors with internal cell retention filters are available for perfusion culture If required it is possible to customize the Cellbag biore actors The following bag sizes are available for Xuri Cell Expansion System W25 e 2L e 10L e 2OL e 22L e 50L 32 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 5 Cellbag bioreactor Illustration of Cellbag bioreactor The illustration shows a general Cellbag bioreactor The configuration of your Cellbag bioreactor may vary from the configuration shown below Component OH bag sensor port
21. OK Cance Cellbag Wainht 4 If preparing a perfusion cultivation calibrate the feed and harvest waste pumps to reach optimal precision Tip For perfusion auto calibration can be enabled which makes this action un necessary Click Calibrate Result The Calibration dialog opens Refer to Calibration instruction on page 202 for instructions on how to calibrate the pump 5 Click OK xuri Cell Expansion System W25 User Manual 29 0646 22 AA 183 5 Operation 5 4 Prepare cultivation 5 4 3 Final checks before cultivation 5 4 3 Final checks before cultivation Follow the instructions below to verify the system before filling the Cellbag bioreactor with medium However the Cellbag must be filled with liquid during this verification Step Action 1 2 Verify that the tray is correctly attached to the rocker platform Verify that the lid is mounted on the tray to protect the optical sensors on the Cellbag bioreactor from excessive light If using both pH and DO sensors verify that the optical fiber cables are not mixed up Mark the cables with the supplied stickers Verify that the pH or DO sensor cables are not positioned betweeen the temperature sensor and the Cellbag bioreactor Verify that the pH and DO sensor adaptors are connected correctly Check if there are visible gas bubbles on the optical sensors See pH reading and DO reading respectively in Section 7 3 Xuri Cell Expansion W25 CBCU on page 2
22. TO Rmeparemern Ome ters E veer OP ferent OT or rN NOD ee Ono tn OPT een NO 260 8 8 7 DH G10 10 22 612 seiis e T rata eerie tetris Tali 265 8 8 8 PR O O AAA A A 268 8 9 9 DG Fogle eis 012 50 e g sasoitan e r e rn her et mn mee Retr pare vren nts 276 cp omit 2 DO CONTO srir iaer INOA AEO TO OONO 279 OO OTO O aa E TAA 284 PPPE EEI A ES IAA E O EE A T E E E AEE sees eas 287 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Page intentionally left blank 1 Introduction 1 Introduction Purpose of this manual Xuri Cell Expansion System W25 User Manual provides you with instructions and infor mation how to run Xuri Cell Expansion System W25 It also includes relevant guidance for practical handling and maintenance of the system units Scope of this document This manual covers Xuri Cell Expansion System W25 including the rocker CBCU pump UNICORN software and accessories In this chapter This chapter includes important user information intended use of the system and lists of important concepts and user documentation This chapter contains the following sections Section See page 1 1 Important user information 8 1 2 Important concepts 10 1 3 User documentation 11 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 7 1 Introduction 1 1 Important user information 1 1 Important user information Read this before operating Xuri Cell Expansion System W25 All users must read the
23. The control parameters of your configuration may vary from the list below Control of Description Temperature The system has a capacity to control temperature between room temperature plus 5 C and 40 C Rockingspeed Rocking speed can be set between 2 and 40 rpm Rockingangle Rocking angle can be set between 2 and 12 degrees Rocking mo Rocking motion can be set between 15 and 100 and is by de tion fault 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 has a capacity to control the gas flow into the cellbag between 0 02 and 1 00 Ipm Media distribu The system has a capacity to control culture medium addition and tion removal Two different modes exist media addition and perfusion pH pH is measured and controlled between pH 6 and pH 8 Three dif ferent modes of pH regulation exist with COs CO base and acid base DO is measured between 0 and 250 and controlled between 0 and 100 air saturation Three different modes of DO regulation exist with O gt speed and O speed CO concentration in gas mix can be controlled between 0 and 15 and measured between 0 and 20 O concentration in gas mix can be measured between 0 and 50 and controlled between 0 and 50 with N and between 21 and 50 with air Xuri Cell Expansion Sy
24. air saturation DO calibration is when the Cellbag bioreactor is flushed with 100 air If you perform the DO calibration with air mixed with 0 10 CO it is possible to calibrate DO down to 90 air saturation 100 air saturation DO calibration must not be performed if N is used instead of air Note CO regulation is slow so it takes some time before the CO concentration reaches the setpoint NOTICE Do not move the rocker during a run since this could damage the scale function and disturb the weight measurement For recommendations on operating conditions refer to Section 4 5 Recommended oper ating conditions on page 141 Follow the instructions below to equilibrate the culture medium Step Action 1 Set the desired rocking speed and angle in Settings Rocking in the Process Picture Start the rocking by clicking the right hand side of the Rocking button Rocking 20 rpm 2 Set the desired gas flow in Settings Gas control Gas flow in the Process Picture Start the gas flow by clicking the right hand side of the Gas flow button 3 If applicable turn on CO mixing by clicking the right hand side of the CO2 button in the Process Picture 4 Click the right hand side of the Temp button to start heating Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 4 Prepare cultivation 5 4 5 Equilibrate the culture medium Step Action 5 When target temperature has been attained do the
25. narrow Use the manual instruction pH control pH control CO2 to set the desired CO range for pH control The automatically se lected control parame ters are not optimal for the actual case Use the manual instruction pH control ad vanced pH control advanced CO2 Set PID pa rameter mode and or Cycle time mode to Manual and enter desired values for PID parameters and cycle time See Manual instructions on page 47 To get an initial suggestion of the PID parameters look at the automatically selected parameters by selecting them as run data under Tools Customize Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 215 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Error symptom OH control using acid or base seems to react too slow or too fast undershooting over shooting Possible cause The molarity of acid or base is not entered correctly Corrective action Enter the correct molarity of the acid and base used See Flow rate on page 125 and Section 5 4 2 Pre culture preparation on page 182 for more infor mation The inner diameter of the tubing used for acid and base is not entered correctly Enter the correct inner diameter of the tubing used for acid and base The weight measured by the system is not corresponding to the weight of the cell cul ture medium Make sure that the rocker scale is tared so that the measured weight corresponds to the weigh
26. remove the protective black bag just prior to use 2 Place the Cellbag bioreactor on a steady surface with the bag sensor ports facing upwards The optical sensor spots have different colors The spot on the pH sensor bag port is white and the spot on the DO sensor port is pink black If both pH and DO sensors are used one fiber cable is needed for each sensor 3 Attach the sensor adapter with the optical lens facing the sensor port by inserting the four pins of the port into the corresponding holes of the adapter Note The sensor adapter can be fastened in any of four orthogonal directions Select the most convenient direction 156 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 2 Prepare pH and DO sensors Step Action 4 Rotate the sensor adapter clockwise to fix the pins on the sensor port to the adapter A distinct click will indicate that the adapter is securely fastened Note When rotating the sensor adapter make sure not to place any force on the fiber cable 5 Place the Cellbag bioreactor on the tray with the optical sensors facing downwards NOTICE Make sure that the optical fiber cables are not placed between the Cellbag bioreactor and the temperature sensor on the tray This could lead to erroneous tem perature reading and control resulting in overheating 6 To keep track of the optical fiber cables mark the cables
27. 0646 22 AA 167 5 Operation 5 1 Prepare the system 5 1 5 Connect gas to system Step Action 2 Connect tubing from the GAS MIX OUT on the CBCU front panel to the inlet gas filter of the Cellbag bioreactor 3 Connect the desired gas source air or N at 1 2 bar 1 0 to 1 5 bar to AIR N2 on the CBCU rear panel 4 If applicable connect the CO gas source at 1 2 bar 1 0 to 1 5 bar to CO2 IN on the CBCU rear panel 168 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 5 Connect gas to system Step Action 5 If applicable connect the O gas source at 1 2 bar 1 0 to 1 5 bar to O2 IN on the CBCU rear panel NOTICE Make sure to keep the inlet pressures within the stated limits 1 0 to 1 5 bar Too high pressure may cause internal tubing to loosen NOTICE An unsteady inlet pressure will affect the speed of the gas flow control and also the gas mix xuri Cell Expansion System W25 User Manual 29 0646 22 AA 169 5 Operation 5 2 Start the system 5 2 Start the system Introduction This section describes how to start the system log on to UNICORN and connect the system to UNICORN Start the system and log on to UNICORN Follow the instructions below to start the system and log on to UNICORN The workstation must have a valid e license For more information about e licenses refer to Operating Instructions Step Action
28. 1 Turn on the client computer 2 Double click the UNICORN icon on the desktop UNICORN 6 3 Result The Log On dialog opens 170 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 2 Start the system Step Action 3 In the Log On dialog e select User Name and e enter Password Note It is also possible to select the Use Windows Authentication checkbox and enter a network ID in the User Name field f Log On UNICORN C Use Windows Authentication UserName Eric vj Domain Aecess Administrators Mi Toup e click OK Result The UNICORN modules the user has chosen under Options are opened EF Evaluation T Administration System Control amp Method Editor User Fale Te Meth Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 171 5 Operation 5 2 Start the system Step Action 4 Switch on the rocker It is blinking green while starting up EE EE HA at Note Pressing the power button while the rocker is on will shut down the system and stop any ongoing run Connect to system Follow the instructions below to connect the system to UNICORN Step Action 1 When the indicator light on the rocker front panel shows a steady green light click the Connect to Systems icon in the System Control module P Result The Connect to Systems dialog opens Connect to Systems Connected systems 1 selected max 3 System name
29. 2 Select the instruction in the block after which you want to add the new in struction Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 5 4 Text instructions Step Action 3 Open the Instruction Box if it is hidden Do the following 1 Set the appropriate breakpoint in the Breakpoint box Note A Breakpoint defines when an instruction will be executed The time set is relative to the start of the block 2 Choose the instruction type and the instruction in the Instructions field For basic help on each instruction select the instruction and press F1 3 Type values for instruction parameters in the Parameters text boxes The allowed range is shown in brackets beside the text box If a scroll bar appears at the right side of the Parameters field additional param eters are available Instruction Box Breakpoint Instructions ja Parameters for Gas flow 0 00 F hour Ait Gas flow setpoint 0 02 1 00 E Ea Var Gas flow Var 0 30 Imin Stat as A Deviation alarm art gas flow Stop gas flow Ott On aoe Sapte Lr Allowed deviation up 0 00 0 20 xit fast fill mode eis Select air source Var 0 05 Z l min ai a a7 Allowed deviation down 0 00 0 20 i z E al ri Var 0 05 l min ih cones eiivanceil Deviation alarm delay 0 3600 DO sensor v 60 sec lt Edit Variable Import Block 4 Click the In
30. 241 8 Reference information 8 3 Client computer specifications 8 53 Client computer specifications The table below describes the recommended client computer specifications for a UNICORN system oper ating with the bioreactor system The operating systems recommended for installation are Windows XP Professional x86 with SP3 installed and Windows 7 Professional x86 and x64 with SP1 installed In the table below these are listed only as Windows XP and Windows 7 Min free disc space UNICORN Client Database Server Worksta tion instal lation E License Server Common Compo nents Min avail able RAM Disc for mat NTFS NTFS NTFS NTFS NTFS OS OS lan guage Windows xP Windows 7 English U S Code 1033 Windows XP Windows 7 Windows Server 2003 2003 R2 Windows Server 2008 2008 R2 English U S Code 1033 Windows xP Windows 7 English U S Code 1033 Windows XP Windows 7 Windows Server 2003 2003 R2 Windows Server 2008 2008 R2 English U S Code 1033 Windows XP Windows 7 Windows Server 2003 2003 R2 Windows Server 2008 2008 R2 English U S Code 1033 Architec ture 242 Intel Dual Core or better Windows xP Windows 7 Intel Dual Core or better Windows XP Windows 7 Windows Server 2003 x86 x64 R2 x86 X64 Windows Server 2008 x64 R2 x64 Intel Dual Core or bet ter Window
31. 5 1 Prepare the system 5 1 1 Select tray and Cellbag bioreactor Step Action 3 Attach the upper part of the tray onto the upper part of the rocker platform and slide the tray downwards into place Note Make sure that the hole s in the tray are aligned with the hole s in the rocker platform 4 Plug the tray connector into the rocker platform connector 154 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 1 Select tray and Cellbag bioreactor Detach tray The tray can be detached from the rocker platform in tilt position and in normal position Tilt position is recommended as described in the instructions below Step Action 1 2 Pull the tray towards you and make sure that the temperature sensors exit the holes of the rocker platform Note If the tray is detached in normal position you need to lift up the tray in the upper edge before pushing it away from you xuri Cell Expansion System W25 User Manual 29 0646 22 AA 155 5 Operation 5 1 Prepare the system 5 1 2 Prepare pH and DO sensors 5 1 2 Prepare pH and DO sensors Instructions Follow the instructions below to connect the sensor adapters to the pH and DO bag sensor ports Step Action 1 Remove the Cellbag bioreactor from its protective cover bag NOTICE Intense light will deteriorate the optical sensors on the Cellbag bioreactor To avoid unnecessary light expo sure
32. Acid tube This parameter will give the acid pump Set pH control yi innerdiam the information needed to estimate the tings acid eter rom that corresponds to the flow re Cellbag quested by the pH control To get a pumps higher accuracy in the flow to rom Acid conversion a calibration of the pump pump can be performed In the pH control A B C case it should however be enough just or D to give the tubing diameter Base mo The molarity of the base used ex Set OH control larity pressed as equivalent of NaOH concen tings base tration mol L If the strength pKb of Cellbag the selected base differs from NaOH pumps try to estimate equivalent molarity or Base see manual for approximate conver pump sion values for other bases The higher A B C molarity of the selected base the less or D amount of base will be added by the pH control and vice versa Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 271 8 Reference information 8 8 Control settings 8 8 8 pH control Setting Description Set from View from PP MI PP RD CD Base tube This parameter will give the base pump Set pH control Y yi innerdiam the information needed to estimate the _ tings base eter rpm that corresponds to the flow re Cellbag quested by the pH control To get a pumps higher accuracy in the flow to rpm Base conversion a calibration of the pump
33. CBCU Corrective action Start media control again 225 7 Troubleshooting 7 4 Xuri Cell Expansion W25 Pump 7 4 Error symptom Pump flow appears to differ from set flow Possible cause Incorrect pump holder position for the tubing used Xuri Cell Expansion W25 Pump Corrective action Check and adjust pump holder position See Sec tion 5 1 4 Prepare the pump on page 161 The setting for tubing inner diameter Is incor rect Reset the tubing inner diameter to correct value Pump needs calibra tion Calibrate the pump See Section 6 1 Calibrations on page 201 Status LED on the pump rear panel is blinking red An internal error has occurred but the pump is still operating Check any warning message and follow the instruc tion If problem persists contact GE Healthcare service personnel Status LED on the pump rear panel shows a steady red light An internal error has occurred and the pump is not operating Check any warning message and follow the instruc tion If problem persists contact GE Healthcare service personnel CAN indicator LED is blinking 226 An internal error has occurred and the CBCU is not operating proper ly Note This is normal the first seconds during power up Contact GE Healthcare service personnel Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 7 Troubleshooting 7 5 Cellbag bioreactor 7 5 Ce
34. Control View C SR System C GS System2 FE System3 EY Se Systema E SS Systems C Ge System C SB System 172 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 2 Start the system Step Action 2 In the Connect to Systems dialog e Select the system e Select Control mode e Click OK Result The Process Picture appears Settings Q Me OTS ek ele ee Temp 19 6 ee ee a bme Ont gt heer Celtep 10t Tip If UNICORN is unable to connect to the selected system see Section 7 6 2 UNICORN System Control on page 232 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 173 5 Operation 5 3 Edit system properties 5 3 Edit system properties Introduction Instruction 174 The system properties of a system can be edited For example if you want to e Change the system setup for example if a CBCU or pump has been added or removed e Change the Instrument Configuration of the system The Instrument Configuration is the system specific control software delivered on a DVD or possible to download from the GE Healthcare homepage The table below describes how to edit the properties of a system Step Action 1 In the Tools menu open the Administration module Click System Properties Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 3 Edit system properties Step Action 2 e Select your system in the System P
35. Information tab contains general information about the result 94 xuri Cell Expansion System W25 User Manual 29 0646 22 AA Documentation tab Method Information 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 3 Run documentation Contents The Method Information tab shows the general Properties of the method e Name e When and by whom the method was created e When and by whom the method was last modified e The system and Instrument Configuration that the method was created for Click the Details button to view comprehensive informa tion about the Instrument Configuration The Method Information tab also contains a listing of all the electronic signatures that have been added to the method under the sub tab Signatures Start Protocol The Start Protocol tab shows the method items that were included in the Start Protocol at the start of the method run System Information The System Information tab shows the system settings during the method run for example e Heating settings e pHand DO sensor settings e Pump setup Calibration The Calibration tab shows a calibration report for the system Run Log The Run Log tab shows selected log entries and feedback Evaluation Log The Evaluation Log tab lists all of the evaluation opera tions that have been performed for the result during all sessions including at the end of the method run The log also shows when the result has been acces
36. PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Gas flow control output data Data Description View from RD CD PP Gas flow measurement The measured gas flow from the CBCU Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable xuri Cell Expansion System W25 User Manual 29 0646 22 AA 255 8 Reference information 8 8 Control settings 8 8 4 CO mix control 8 8 4 CO mix control CO mix control settings The following table shows general settings used for CO mix control Setting Description Set from View from PP RD D CO2 Mix set The gas mixer target CO concen Settings Gas CO2 mix point tration setpoint control CO2 Deviation Select On to activate the devia CO2 mix alarm tion alarm and Off to deactivate it Allowed devia A parameter that defines how Settings Gas CO2 mix tion up much above the setpoint the control C02 CO concentration to the Cellbag is allowed to be without trigger ing the deviation alarm Allowed devia A parameter that defines how Settings Gas CO2 mix tion down much below the setpoint the CO control CO2 concentration to the Cellbag is allowed to be without triggering the deviation alarm Deviation A parameter that defines for CO2 mix alarm delay how long the CO concentration to the Cellbag can
37. UNICORN and System Log provides the system administrator with records of usage and activity System Properties is used to define the system and edit system properties Database Management is used for maintenance of the database Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 41 3 The UNICORN software 3 2 System control 3 2 System control Introduction The System Control module is used to start view and control a manual or a method run System Control panes As illustrated below two tabs are available in the System Control module by default The Process Picture tab allows manual interactions with the system and provides feedback on run parameters Data is illustrated as curves in the Chart tab during the entire run Refer to Section 5 5 Perform cultivation on page 191 for information on how to perform a run and to Section 5 5 2 Monitor and control the cultivation on page 193 for information on how to work with the Process Picture Tip To get more information than what is shown in the Process Picture select View Run Data to open the Run Data pane which presents current data in numerical values SS EE onl ty ie F Ta 42 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 2 System control Actions in the Process Picture pane It is possible to interact with the Process Picture pane in the following ways If you wan
38. a volume of 1 liter 3 When the cells reach 3 x 10 cells mL transfer using a tube fuser to a 20 L Cellbag bioreactor containing 2 liters of culture medium 4 When the cells in the 20 L Cellbag bioreactor reach 3 x 10 cells mL add culture medium up to a volume of 10 liters and continue cultivation Culture medium exchange Follow the instructions below to exchange culture medium in the Cellbag bioreactor for example when working with microcarriers 196 Note To avoid oxygen depletion the culture medium exchange should be performed in less than one hour Step Action 1 Stop the gas flow by clicking the right hand side of the Gas flow button in the Process Picture 2 Stop the rocking by clicking the right hand side of the Rocking button in the Process Picture 3 Clamp off the inlet and outlet gas filters 4 Select Manual Execute Manual Instructions and send the instruction Rocker Prepare for tilt 5 Grab the textured grip area on each side of the tray and pull the tray up wards and against you to position the tray into tilt position 6 Allow particulates including cells and microcarriers if applicable 10 to 15 minutes to settle 7 Connect tubing to the harvest port on the Cellbag bioreactor The other end of the tubing should be connected to a sterile harvest vessel 8 Using a pump or gravity flow remove the desired amount of supernatant culture liquid by manually manipulating the flexible wall of the Cellbag b
39. arse eee 166 D E 15 gt e EA nee eres enn eee 170 5 3 Editsystem properties sapecssstssscvscussscovecoscestorsseravsrbasscdsasssuchysscssbedsbess son uvasbsqvtessesszeucheossslvebossveastestsiiees 174 A Prepare Cutao E E ree omen etn E 176 5 4 1 O TU a E E L7 5 4 2 Pre cult re preparOtiOl sssini rris rarene AEREA tear ecaeveveee 182 5 43 FING CHECKS before cultivation sesssenssinensiasniunsknniisainani 184 5 4 4 Adalionmor culture mediUN suais re e AE 185 5 4 5 Equilibrate the culture medium ssssssssssssssssssiressssssssssssssssssrrrrrssssssssssssssssrrrrrrresessssssssss 188 Do PACU ON ekaa oE SEE EE E AA 191 5 5 1 KOCU I acne EE EAEE ETE AEAN TE E 192 552 Monitor and control the cultivation sscscscsscctsaccossivacsseusvaxsshsnnsstonsenstSausensistesdracscevavevecsstidnavsecis 193 5 5 3 FNG CUNO 16 6 sasar ir E AA ATAOE 198 O i U2 18 gs ere ere ae rn rt E er Re eee een nner emer eee 200 Sm O OS geen tee antec nT et nme ane fe acevo en nT ENE NOT Cty FO en Ue oan 201 Ge PUPE GUO le eee ween eo eee nn OnnnT tea mere vere a on EOD Sera rere ne err 203 ce EA geaeee en neon ere mn De oO E On Ena E E en E eee 205 T Troubleshooting ene eee enon nance aE 206 Tok UII CS I Expansion Systemi W2 agate cceccescsp cies consstesseatvsewriasnciest deadevtesdicvedsonpdebeadeehssetlatwcie 207 V2 RUG Cell Expansion System W25 TOCKE assisia iE Ea iaia 208 T3 AUCE Expansion WAS Cl Gt Dl aanren aR nmnEre vet PnNEEETEE TE ree eC ve
40. base ctrl output C 020 Total acid C 021 Total base V 022 Run Log V 023 DO ia 024 DO setpoint C 025 DO amplitude 3 Select the checkboxes of the curves you want to show in the chart and then click OK Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 2 System control System settings Each installed instrument has a set of default parameter values called system settings The System Settings dialog in System Control is used to view and edit the system setting for the currently selected instrument before the run is started Follow the instructions below to change the System Settings All available System Settings are described in System Settings Step Action il In the System Control module select System Settings Result The System Settings dialog opens with the Instructions displayed See examples of different menus below Instructions Heating pH sensor DO sensor Pump setup Auto start 2 Select the instruction to edit from the list Click the symbol to show the in structions for each category The instructions in each category differ depend ing on the instrument configuration 3 Select settings and choose parameter values for the selected instruction Click OK 4 To return to the default values defined in the instrument configuration click Set Parameters To Strategy Default Values Manual instructions It is possible to manua
41. be outside the allowed deviation limits before the deviation alarm is triggered Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable 256 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 4 CO mix control CO mix control output data Data Description View from RD CD Pp CO2 measurement The measured CO concentration from the CBCU Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 257 8 Reference information 8 8 Control settings 8 8 5 O mix control 8 8 5 O mix control O mix control settings The following table shows general settings used for O mix control Setting Description View from PP RD D 02 Mix set The gas mixer target O concen Settings Gas point tration setpoint control 02 Deviation Select On to activate the devia alarm tion alarm and Off to deactivate it Allowed devia A parameter that defines how Settings Gas tion up much above the setpoint the O control 02 concentration to the Cellbag is allowed to be without triggering the deviation alarm Allowed devia A parameter that defines how Settings Gas tion down much b
42. by time the user when DO control is off and by DO control when the DO control is on DO measurement The latest measured DO value DO amplitude A measure of the strength of the responsive DO signal received by the DO monitor DO phase The phase shift between a reference signal and a signal representative for the oxygen content at the sensor Used to calculate the actual DO DO temp comp Temperature used for temperature compensation of the DO value DO LED current A measure of the intensity of the LED used to illuminate the DO sensor Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable 278 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 10 DO control 8 8 10 DO control Introduction The following tables describe settings and parameters that can be used for DO control General settings The following table shows general settings used for DO control Setting Description Set from View from Select Speed for using only rocking speed as actuator Se lect O2 for using only O as actu ator Select O2 Speed for a combination of O and rocking speed as actuators where the control always starts in O mode Settings DO DO control general dad gt Oe D DO setpoint The DO target value Settings DO DO control genera
43. by clicking the right hand side of the Gas flow button in the Process Picture Result The gas flow is maximized to 3 L min for 20 minutes or until Fast fill is manually turned off The cellbag is inflated Stop the gas flow when the cellbag has been fully inflated by clicking the right hand side of the Gas flow button xuri Cell Expansion System W25 User Manual 29 0646 22 AA 149 5 Operation 5 Operation In this chapter This chapter describes how to operate Xuri Cell Expansion System W25 This chapter contains the following sections Section See page 5 1 Prepare the system 151 5 2 Start the system 170 5 3 Edit system properties 174 5 4 Prepare cultivation 176 5 5 Perform cultivation 191 150 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 Prepare the system In this section This section describes how to prepare the bioreactor system for cell cultivation For illus trations and descriptions of the system refer to Chapter 2 System description on page 12 This section contains the following subsections Section See page 5 1 1 Select tray and Cellbag bioreactor 152 5 1 2 Prepare pH and DO sensors 156 5 1 3 Attach the Cellbag bioreactor 159 5 1 4 Prepare the pump 161 5 1 5 Connect gas to system 166 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 151 5 Operation 5 1 Prepare the system 5 1 1 Select tray and Cellbag bioreactor 5 1 1 Select
44. content in the Cellbag bioreactor and the molarity of the acid and base The flow rate is further discussed below PI parameters and cycle time The automatically computed Pl parameters are static in the Acid Base control mode They can be seen as default parameters The same holds for the automatic cycle time These automatic values should be good enough for most cases and no tuning is expected to be needed In the acid base controller the part of the output is only updated when the pH mea surement is close to the setpoint When elaborating with a manual Il parameter remember that increasing the I parameter will reduce the integrating effect and vice versa Tuning of PI parameters and cycle time If tuning of the PI parameters or cycle time is needed follow these steps Step Action 1 Open Tools Customize and select the tab Run Data Groups Select the Auto pH acid base P and Auto pH acid base I and click OK Result The automatic PI parameters are shown as Run Data 2 Open the manual instruction pH control advanced pH control advanced acid base and select Manual PID parameter mode and or Cycle time mode and enter the desired values Click Execute 124 xuri Cell Expansion System W25 User Manual 29 0646 22 AA Flow rate 4 System control description 4 2 pH control 4 2 2 Acid Base control mode Xuri Cell Expansion System W25 automatically computes flow rates for acid and base There is no need for calibration
45. delays O2 Speed control scheme 132 The O02 Speed control scheme is combining the O2 and Speed control modes It always starts in 02 mode and can then shift back and forth between O2 and Speed mode as long as the control is on To avoid unnecessary toggling between O2 and Speed mode the transitions between the modes are delayed This delay is called transition delay time The condition for transition must be fulfilled for a certain time before the transition takes place The transition delay times are set automatically and individually by the DO control The values can be overridden with manual values using the manual instruction DO control advanced DO control transition delays 02 mode Condition Condition DO reading gt DO set point DO reading lt DO set point AND AND Rocking speed set point at 02 set point at maximum minimum fullfilled for the fullfilled for the O2 to speed transition delay time Speed Speed to O2 transition delay time mode If the DO O controller is at its maximum O setpoint 50 and the DO reading is below the DO setpoint a time counter is started When the time counter reaches the set 02 to speed transition delay time the transition to Speed mode takes place If the DO speed controller is at its minimum rocking speed setpoint and the DO reading is above the DO setpoint a time counter is started When the time counter reaches the Speed to 02 transition delay time the transition to O mode takes pla
46. describes this Step Action ii e Inserta vertical marker where you want to take the Snapshot e Right click and choose Snapshot from the shortcut menu Result The Snapshot dialog opens z 3 yea JNN Ei oe eal SELLS mn ba on Len m a EREL Fy e gt i O a ca a a ap ap ap ap z o e Saa Pia Cista 2 e Click the Save button to save the Snapshot Result The Save As dialog opens and you can save the Snapshot as a text file or e Click the Print button to print the Snapshot Result The Print dialog opens and you can print the Snapshot using the selected printer Note Snapshots taken during the method run are saved directly to the result and can be accessed by clicking the Snapshot sub tab in the Result Information tab of the Documentation dialog 3 Click the Close button to close the Snapshot dialog Note The snaphot will record only the values of curves that are displayed Curves that are filtered will not be recorded 92 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 3 Run documentation 3 4 3 Run documentation Introduction The full documentation for a run is stored in the result This section contains e an instruction how to view and print the run documentation e alist and short descriptions of the contents of the run documentation e aninstruction how to save the text instructions from a method
47. devi ation alarm is triggered Heating Heat enabling 252 Note When set to Enabled the heat ing will be on when the rocking is on When set to Disabled the heating will be off System set tings Heat ing Heater en abling In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 2 Heating control Heating control output data Doh Ke Description View from RD CD Pp Temperature measurement The measured Cellbag temperature Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 253 8 Reference information 8 8 Control settings 8 8 3 Gas flow control 8 8 3 Gas flow control Gas flow control settings The following table shows general settings used for gas flow control Setting Gas flow set point Description The gas flow controller setpoint to the Cellbag Set from PP Settings Gas control Gas flow MI Gas flow View from D Y Y Deviation alarm Select On to activate the devia tion alarm and Off to deactivate it Gas flow Allowed devia tion up
48. feed pump is run ning The weight is below the setpoint when the me dia control is started None This is normal The harvest waste pump will start when the setpoint is reached Auto calibration is en abled and the feed pump is calibrating None This is normal The measured weight is more than 10 be low the weight setpoint None This is normal The harvest waste pump will start when the setpoint is reached Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 223 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Error symptom In Perfusion mode only the harvest waste pump is running Possible cause The weight is above the setpoint when the me dia control is started Corrective action None This is normal The feed pump will start when the setpoint is reached Auto calibration is en abled and the har vest waste pump is calibrating None This is normal The measured weight is more than 10 above the weight set point None This is normal The feed pump will start when the setpoint is reached In Perfusion mode no pump is running Auto calibration is en abled and pumps are stopped for the auto calibration algorithm to measure the weight None This is normal Auto calibration is set to On but no auto cali bration takes place The weight setpoint has not yet been reached Wait for the setpoint to be reached then the auto
49. meer 212 LA Kur CelExpansion W25 PUMO eresien od eS 226 T739 06 2 DIOredCtOr soseri anean ane Ege NE A E E 227 Yc SONWare 8 010 a 9 cae eee enn eee mene nec ene rn eee EER 229 7 6 1 Troubleshooting Method editor seecucscicsicssssicinenestiaicicssosiensccasedetnuniitenctsaeneiartenenies 230 7 6 2 UNICORN System CONTO sensusnnnni enean 252 705 Troubleshooting EVQIIOUON ccczc ctccesexcresedesscenaictascccasstdcies ukstecacteizsteteetearnnnniccaiedeaas 255 8 ROTSPEMC A informatio secies ne a 236 81 Gyslem SpeciNcatONE csser sneen nee AEE aA nE aE Oe ai 237 8 2 Component specifications s sssesssesssssssessesssssssssssssssssissrissiririrsrrerrrerrerrnnnrnssnnnnnnnnnnnnnnnnnnnnnnnnsnnntennnn 238 4 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 3 8 4 8 5 8 6 8 7 8 8 Table of Contents Glient computer SHPECICOIUONE comets anne neem ene nen een rn veer nn rere Pe 242 xuri Cell Expansion W25 CBCU Semi wetted materials oo cccsessssesssecsseesseesseesseesseeesseen 244 Cremical tesis 00 9 ee eae eee ne eee a nT ae ce ee 245 Tubing and Connectors seinare a EE nETIE mn TREO ee Eee 246 PUM A OVA ae EEE E E E 247 CTITEL e E E E E 248 8 8 1 POT O r EEE E A EEE 249 8 8 2 Hegting CONTO sirnnennnoien r re n A E ENEA OEA resrrer err 252 8 8 3 OS OW CO Gidea a oe aennanidacustata anaemia RE 254 8 8 4 BOP io LE Sennen ee enn mr EANN E E 256 8 8 5 OP E 2 6 PA AA TE AA eee TSC eae Tera ce 258 8 8 6 it a Glo
50. min 0 00 Heating 37 0 Heating setpoint C On 0 5 C 1 0 C 60 sec 0 00 Gas flow 0 30 Gasflow setpoint l min On 0 05 l min 0 05 l min 60 sec 0 00 CO2 mix 5 0 CO2 setpoint On 1 0 1 0 60 sec Set pH reading cycle time 120 pH reading cycle time sec 0 00 Start rocking 200 00 Duration End Block Breakpoint Instructions es a procera Parameters for Set pH reading cycle time 00 5 hour fugit Reading cycle time 30 1000 i 5 g ap abo VAR 120 sec Var Gas flow CO2 mix 02 mix pH sensor pH sensor calibration values Set pH reading cycle time Start pH reading Stop pH reading pH control pH control advanced DO sensor DO control H DO control advanced v lt Edit Variable Import Block 2 Click the VAR button for the appropriate variable Result The Edit Variable dialog opens Edit Variable Vanable name 1H reading cycle time Visible in details only 3 Do one or several of the following as appropriate e Type in a new name in the Variable name field e Check the Visible in details only if the variable should be a detailed variable Uncheck the box to set it to a normal variable e Click Clear to delete the variable 70 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 5 4 Text
51. on the feed and harvest waste pumps will be calibrated automatically by the system when the weight SP is reached then with the time interval given by Auto calibra tion cycle If an extra auto cali bration is wanted select Off and then On again Settings Me dia control Media control general Auto calibra tion cycle If auto calibration is on this pa rameter defines the time interval between the auto calibrations Media control general Feed tube in ner diameter The inner diameter of the feed tubing Settings Cell bag pumps Feed pump A B C or D Feed Harvest tube inner diame ter The inner diameter of the har vest waste tubing Settings Cell bag pumps Har vest Waste pump A B C or D Har vest Waste 1 This run data is not specifically connected to the feed or harvest waste role Instead this run data is connected to a specific pump head Look at the run data from the pump head of interest Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 285 8 Reference information 8 8 Control settings 8 8 11 Media control Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Media control output data Description View from Pp Momentary weight set In Media addition mode the weight setpoint is ramped point towards the
52. on dialog is inactive and a Verify that no UNICORN window or module is opened password cannot be entered Access to UNICORN functions Problem description The Execute manual instruction menu command in the System Control module is gray This means that you can establish a connection but cannot control the system Log off from Windows and log on again Solution e Check that no other user has a control mode connection e Check that you have access rights to control the system manually The help viewer cannot be opened using help but tons or the F1 key 1 Open the MadCap help viewer from the Win dows desktop icon This is described in The help viewer application in UNICORN Administration and Technical manual 2 Try the help button or F1 key again The Microsoft Office Document Image Writer causes UNICORN to terminate This writer application will not work Choose an other option for example a PDF writer application A user without access to the function Method End may still end a method using a Timer instruction Access to the Timer instruction must also be dis abled if users are not allowed access to the Method End function 232 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Problem description A manual run is started A method run is then started and a start protocol opens Before the start protocol is finished an alarm is caused by the 7 Troubleshooting 7 6 Software pro
53. run as anew method View and print the run documentation The table below describes how to view and print the run documentation Step Action 1 Open a result in the Evaluation module e Choose View Documentation or e Click the view Documentation icon z lt ia Result The Documentation dialog opens See further information about the tabs and contents below 5 e Click the Print button Result The Print dialog opens Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 93 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 3 Run documentation Step Action 4 e Select the documentation items you want to print Note Items that do not contain any information cannot be selected If you select a group heading e g Result information all sub headings that contain information will automatically be selected Printer Adobe PDF Items Result information Properties Name and location C System information Calibration C Evaluation log Bun log 5 Click the OK button to print the selected items The documentation tabs The table below describes the contents of the Run Documentation tabs Refer to the online help for more information about each tab Note Some of the items listed below may be excluded from the Run Documentation Tabs will be shown only for items that have been included in the method Documentation tab Contents Result Information The Result
54. the Cellbag bioreactor Note Pump calibration is normally not needed xuri Cell Expansion System W25 User Manual 29 0646 22 AA 203 6 Maintenance 6 2 Pump calibration Calibration of feed and harvest waste pumps The feed and harvest waste pumps are calibrated at the flow entered as the media control flow rate setpoint e Make sure that the flow rate set point is entered correctly before calibrating the feed or harvest waste pump Media addition mode No calibration is needed since the correct flow is ensured by a transitional weight setpoint see Section 4 4 1 Media addition on page 134 Perfusion mode The pumps are calibrated automatically by the system However when running at a low flow rate below 3 43 L day manual calibration is needed The auto calibration is then unavailable see Section 4 4 2 Perfusion on page 135 Calibration factor After calibrating a pump manually or using auto calibration the calibration factor is recomputed The calibration factor is the relation between the expected flow to rom conversion for the selected tube diameter and the result of the calibration For feed and harvest waste pumps this value can be observed in the curve data chart Warning and rejection limits The pumps will give a warning if the given calibration volume deviates more than 30 to 10 from the expected value The calibration will be rejected if the given calibration volume deviates more than 60 to 30 f
55. the instruction of interest in the Instruction box in the Text instruction pane press the F1 key In the System Control module e select Manual Execute Manual Instructions expand a heading and select the instruction of interest press the F1 key or click the Help icon in the dialog Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 39 3 The UNICORN software 5 The UNICORN software In this chapter This chapter gives an overview of how to work with the four UNICORN modules Detailed information about the UNICORN software is available in the UNICORN user documentation and in the UNICORN Online Help This chapter contains the following sections 40 Section See page 3 1 Administration 41 3 2 System control 42 3 3 Methods in UNICORN 49 3 4 Evaluation in UNICORN 86 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 1 Administration 3 1 Administration Introduction The Administration module is used to administer all functions of the UNICORN software Refer to UNICORN Administration and Technical manual for more information Icons in the Administration module The table below shows the Administration module icons Function User Setup is used to administer the user access to UNICORN Access Groups and Network Users is used to administer access groups and network users E mail Setup is used to set up an e mail account for automated system messages
56. the rocker The illustration below shows the front view of the rocker Description Rocker platform Temperature sensors Rocker base Power switch Location of adjustable foot Power switch The power switch indicates the status of the rocker according to the list below Light indicator Description No light The power is off Green blinking light The rocker is starting up Green steady light The power is on and the rocker is operational Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 17 2 System description 2 2 Xuri Cell Expansion System W25 rocker 18 Light indicator Description Red blinking light The rocker failed to connect to 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 when viewed from the front It is used to evenly distribute weight over the four rocker feet To adjust the foot remove the side plate carefully lift the front right corner of the rocker and turn the foot by hand In the image below the rocker is shown without a side plate in order to display the ad justable foot Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 2 Xuri Cell Expansion System W25 rocker Rear view of the rocker The illustration below shows the rear panel of the rocker Description 15 pin D sub connector Filter h
57. the rocking speed should not be set higher than 30 rpm Rocking motion Gas flow 142 The rocking motion sets how large part of the rocking cycle that has a sinusoidal angular velocity It is by default set to 30 It is possible to change the rocking motion between 15 and 100 where 15 gives a uniform motion with almost constant angular veloc ity throughout the movement and 100 gives a sinusoidal motion The lower percentage will give slightly higher oxygen transfer rate and mixing capacity whereas smoother rocking profiles can reduce shear forces especially important when cultivating adherent cells on microcarriers The gas flow rate has little effect on oxygen transfer and can therefore be kept constant throughout the entire run When setting the gas flow rate the following should be taken into consideration e Ahigh gas flow can cause unnecessary evaporation e Allow gas flow can slow down pH and DO control when using CO and O gas e Atoo low gas flow can lead to insufficient inflation of the Cellbag bioreactor Refer to Gas flow table for recommendations on suitable gas flow rates Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 5 Recommended operating conditions Recommendations to reduce foaming To reduce foaming the following should be taken into consideration e Higher agitation rates can contribute to excessive foaming This can be reduced by adding antifoam
58. to Section 5 1 5 Connect gas to system on page 166 In the Process Picture click the Compressed Air button to select compressed air or the N2 button to select N gt 6 Start the gas flow by clicking the right hand side of the Gas flow button 7 Enter a gas flow setpoint for example 0 2 I min in Settings Gas control Gas flow Setpoint Click OK 8 Enter a O setpoint for example 30 in Settings Gas control 02 Setpoint Click OK 9 Wait for a few minutes and verify that the Process Picture shows a correct measurement If the gas flow appears not to function correctly refer to Section 7 3 Xuri Cell Expansion W25 CBCU on page 212 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 6 System verification Verification of fast fill function Follow these instructions to make sure that the Fast fill functions properly Step Action 1 g Switch on the client computer Switch on the rocker and wait for the system to initialize Start UNICORN and connect to system Make sure that the desired gas supplies are connected to the system Refer to Section 5 1 5 Connect gas to system on page 166 Open Settings Gas control Gas flow from the Process Picture in System Control Set Fast fill to On O2 Setpoint 1 00 l min c z Alert i 0 054 0 05 Fast fill Enable Disable Cellbag pumps OK Cancel Cellbag Weight Rocking Gas flow Start the gas flow
59. tray and Cellbag bioreactor Size selection guide Select Cellbag bioreactor size and corresponding tray depending on experiment demands and system configuration see table below Cell culture volumes L Cellbag bioreactor size L Tray 10 Tray 20 Tray 10 Tray 20 Tray 20 Tray 50 Tray 50 Note Depending on application and configuration it might be possible to cultivate below the recommended minimum volume However it is highly recommended to stay above this volume for applications that require high agitation and pH and DO control The temperature pH and DO sensors need to be submerged in liquid throughout the complete rocking cycle to function correctly Tip To achieve good temperature control at low liquid volumes it is critical to keep the lowest angle possible that is sufficient to attain adequate mixing and aer ation 152 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 1 Select tray and Cellbag bioreactor Attach tray The tray can be attached to the rocker platform in tilt position and in normal position Tilt position is recommended as described in the instructions below Step Action 1 Tilt the rocker platform by pulling the upper ledge towards you aN m L j ne a 2 Lift the tray into the same angle as the rocker platform xuri Cell Expansion System W25 User Manual 29 0646 22 AA 153 5 Operation
60. w BU lt sec gt Edit anable Import Block xuri Cell Expansion System W25 User Manual 29 0646 22 AA 67 3 The UNICORN software 3 3 Methods in UNICORN 3 3 4 Text instructions Edit variables Editing a variable includes renaming and deleting the variable and choosing whether the variable should be a detailed variable or not Edit a variable using the Edit variable button The table below describes how to edit a variable using the Edit Variable button Step Action 1 e Inthe Instruction Box click Edit Variable Breakpoint i Instructions a SN Parameters for Heating 0 00 5 hour HEN sij Temperature setpoint 15 0 40 0 Heating ripe Vat Heating 37 5 C Sdi lala Deviation alarm isable heating Gas flow Off On m CO2 mix Allowed deviation up 0 2 5 0 O2 mix 5 _ mE pH sensor Var f 0 5 2 C ot coe aruge Allowed deviation down 0 2 5 0 pH control advance ML DO sensor 7 t 1 0 2 C m DO control Deviation alarm delay 0 3600 DO control advanced i a Media control Var 60 7 sec lt gt Edit Variable Import Block e Alternatively select the Phase Properties tab to display the phase vari ables select the variable and click Edit Variable Result The Edit Variable dialog opens displaying all variables if opened from the Text Instructions pane or the phase variables if opened from t
61. 12 for advise on how to remove any bubbles Note Such bubbles may be difficult to observe Verify sure that the filter heater is connected Verify that the Cellbag bioreactor is firmly inflated and secured to the tray The Cellbag bioreactor should be taut but not creased Verify that gas flow is released through the outlet pressure relief valve This may be done by attaching a piece of tubing with one end submerged in a beaker of water to the pressure relief valve Formation of gas bubbles in the water will confirm its function 184 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 4 Prepare cultivation 5 4 4 Addition of culture medium 5 4 4 Addition of culture medium Important It is important to tare the scale with all the equipment on the tray such as lid Cellbag bioreactor and filter heater The measured weight should for optimal control be the same as the weight of the culture The weight measurement is used as input for the regulation of temperature pH and media control Make sure that the rocker stands on a horizontal surface and that the tray is in a hori zontal position NOTICE The rocker has a scale function load cells and should therefore be handled with extra care The rocker shall not be placed on any parts that may move Avoid exposing the load cells to lateral forces such as pushing it sideways on the bench which may cause distur bances in the measurement or damage the loa
62. 2 Method creation The table below describes how to create a new method using a predefined method as template Step Action 1 In the Method Editor click the Create a new method icon in the Toolbar or select File New Method Mew Method Ctrl M Mew Method Queue Open Chrl 0 Close Result The New Method dialog opens Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 53 3 The UNICORN software 3 3 Methods in UNICORN 3 3 2 Method creation Step Action New Method Create a new method by using the Predefined Method WAVE 25 Template Empty Method Method Description This predefined method may be used to create methods for a Ready T oProcess WAVE 25 system 2 In the New Method dialog e select a System e select a Predefined Method e click OK Result The Method Outline pane shows the mandatory Method Settings phase for the chosen method and the Text Instructions pane shows all the instructions that defines the method The Phase Properties pane shows the default settings for the currently highlighted phase 54 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 2 Method creation Empty method The table below describes how to create a new empty method Step Action 1 In the Method Editor e click the Create a new method icon in the Toolbar or e select File New Method
63. 9 0646 22 AA 63 3 The UNICORN software 3 3 Methods in UNICORN 3 3 4 Text instructions 5 5 4 Introduction Text instructions When a user defined phase in the Method Editor is selected the corresponding phase block is selected in Text Instructions when changing to the Text Instructions tab Changes made in the Phase Properties pane are automatically updated in the Text In structions pane Text editing a method Adding editing or deleting any blocks or instructions in a phase in the Text Instructions area means text editing of the method When a method has been text edited one or several of the phases displayed in the Method Editor window are affected depending on the type of editing performed The letter T next to the phase name in the Method Editor window indicates that the phase has been text edited Help texts for the instructions It is possible to display help texts for the instructions in the Instruction Box The table below describes how to display the help text for an instruction Step Action 1 In the Instruction Box select the appropriate instruction for which to display help text 2 Press F1 on the keyboard Result A dialog with help text for the selected instruction will be displayed Insert a new instruction 64 The table below describes how to insert a new text instruction in the Text Instructions area Step Action 1 Select a block and display the instructions within the block
64. A parameter that defines how much above the setpoint the gas flow reading is allowed to be without triggering the deviation alarm Settings Gas control Gas flow Gas flow Allowed devia tion down A parameter that defines how much below the setpoint the gas flow reading is allowed to be without triggering the deviation alarm Settings Gas control Gas flow Gas flow Deviation alarm delay A parameter that defines for how long the gas flow reading can be outside the allowed devi ation limits before the deviation alarm is triggered Gas flow Air source If set to Compressed air com pressed air shall be connected to the inlet on the back of the CBCU If set to N2 pressurized nitrogen shall be connected to the inlet on the back of the CBCU Air source 254 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 3 Gas flow control Setting Description Set from View from daa gt Oe Auto start By setting this parameter to Not System set mode resume activity gas mixing will tings Auto not start automatically at start start CBCU up of the system at any time When the parameter is set to Resume activity the gas mixing will start automatically at start up if it was on when the system was shut down without first go ing to END Note In the table the following abbreviations are used
65. Action 1 Open a result file 2 e Select File Report or e Click the Report icon A Result The Generate Report dialog opens 3 e Select a report format to edit e Click the Edit button Result The Customize Report window opens 4 e Make the necessary changes to the report format Note Once you have made a change the title bar of the window will add the word Modified to the report format name a e Select File Save or e Click the Save icon Result The edited format is saved Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 111 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 6 Edit an existing report format Step Action 6 To return to the Evaluation module window e Select File Exit or e Click the Exit button i e Click the Close button to close the Generate Report dialog or e Select another report format to edit Note See Section 3 4 5 Create a new report format on page 100 for instructions about how to add or edit report items You can also select File Save As in the Cus tomize Report window to save the edited format under another name and keep the original report format unchanged 112 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 System control description In this chapter This chapter describes pH DO and media control Moreover it contains recommended operating conditions and verification procedures for th
66. DO sensors are used check that the sensor cables are not mixed up If so correct the connections of the pH and DO sensors Note that it may take one reading cycle before correct values are shown Check that the correct calibration values as stated on the Cellbag have been entered See Sec tion 5 4 4 Addition of culture medium on page 185 for more information Note The value CpHO may have changed after recalibra tion This is normal Temperature of the cul Turn off the pH reading until the culture medium ture medium has not is equilibrated reached process tem perature OH inside the Cellbag If possible adjust the Cellbag bioreactor conditions bioreactor is outside to a pH within the working range the readable range ap pr pH 5 9 NOTE It is a risk that the pH sensor has been damaged especially if the pH has been above pH 10 218 xuri Cell Expansion System W25 User Manual 29 0646 22 AA Error symptom Initial pH reading is un stable and or deviates considerably i e more than approximately 0 5 OH units from an offline measurement Possible cause A gas bubble may be trapped on the pH sen SOT 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Corrective action Tap the pH sensor from the underside to remove the bubble This may require forceful manipulation and the Cellbag bioreactor may need to be partly disconnected from the tray Note The gas bubble may be very difficult to see Fluc
67. ICORN help 38 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 6 UNICORN software overview 2 6 1 General UNICORN operation 2 6 1 General UNICORN operation UNICORN modules overview UNICORN consists of four modules System Control Evaluation Administration and Method Editor The main functions of each module are described in the table below Module Main functions Method Editor Create and edit methods System Control Start view and control runs Evaluation Open results evaluate runs and create reports Administration Perform user and system setup system log and database administration Enter a UNICORN module To enter a module e click the Taskbar button of the module of interest S administration EF Evaluation g4 System Control ie Method Editor or e choose the module of interest in the Tools menu in any of the other software modules The illustration below shows the Tools menu of the Evaluation module Administration Method Editor System Control LUStomlzs ss Options Change Passwords xuri Cell Expansion System W25 User Manual 29 0646 22 AA 37 2 System description 2 6 UNICORN software overview 2 6 2 The UNICORN help 2 6 2 The UNICORN help Access the help utility A comprehensive help utility is included in the UNICORN software The table below de scribes how to access the different parts of the help utility If y
68. K Result The rocking speed changes to the set value 2 Try some different setpoints of the rocking speed and make sure that the rocking speed changes 144 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 6 System verification Step Action 6 Follow these instructions to make sure that the rocking angle functions properly 1 While the tray is rocking enter the desired rocking angle in Settings Rock ing Angle in the Process Picture Click OK Result The rocking angle changes to the set value 2 Try some different setpoints of the rocking angle and make sure that the rocking angle changes 7 Follow these instructions to make sure that the rocking motion functions properly 1 While the tray is rocking select Manual Execute Manual Instructions and open the instruction Rocker Set rocking motion Enter the desired rocking motion Click OK Result The rocking motion changes to the set value Note The rocking motion sets how big part of the rocking cycle that has a sinu soidal angular velocity 2 Try some different setpoints of the rocking motion and make sure that the motion changes 8 If the rocking speed rocking angle or rocking motion appear not to function correctly refer to Section 7 2 Xuri Cell Expansion System W25 rocker on page 208 9 Stop the rocking by clicking the right hand side of the Rocking button Verification of temperature control Follow the
69. P RD CD pH measurement cycle time The actual measurement cycle time This value is set by the user when pH control is off and by pH control when the pH control is on pH measurement The latest measured pH value pH amplitude A measure of the strength of the responsive pH signal received by the pH monitor pH phase The phase shift between a reference signal and a signal representative for the pH at the sensor Used to calculate the actual pH pH temp comp Temperature used for temperature compensation of the pH value pH LED current Note A measure of the intensity of the LED used to illuminate the pH sensor In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 267 8 Reference information 8 8 Control settings 8 8 8 pH control 8 8 8 Introduction The following tables describe settings and parameters that can be used for pH control pH control General settings The following table shows general settings used for pH control Setting Description Set from View from PP MI ed ey D E D Control Select CO2 Base for a combina Settings pH pH control Y scheme tion of CO and base as actua general tors where the control always starts in CO2 mode Select Acid Base for a comb
70. P RD CD CO2 Base When Auto is selected the system OH control A hand transition de will automatically set the transition transition symbol is lay mode delay time The automatically comput delays shown if ed transition delay times can be dis set to played as run data When Manual is Manual selected the system will use the manual transition delay times set by this instruction Manual CO2 Thetransition delay time from CO to PH control to base delay base control used if CO2 Base transi transition tion delay mode is set to Manual delays Manual base The transition delay time from base OH control to CO2 delay to CO control used if CO2 Base transition transition delay mode is set to Man delays ual Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Transition delay output data Description View from d a 1 D EEEE D Auto CO2 to base transition The CO to base transition delay time used if pH delay CO2 base transition delay mode is set to Auto Auto base to CO2 transition The base to CO transition delay time used if pH delay CO2 base transition delay mode is set to Auto Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable xuri Cell Expansion System W25 U
71. Release the mouse button Result A Setup dialog opens The dialog is specific to the type of item that you want to insert For more information about the setup dialogs for each object refer to the online help 106 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Step Action 3 e Select the desired Settings for example Start on new page and click OK Result The object is inserted onto the page Note e Ifyou want to edit an object later double click the object box e The size of the object in Edit mode does not always correspond to the size in the printed report Click Preview in the toolbar for a print preview Move and resize objects freely The table below describes how to select move and resize objects freely If you want to select a single object e click the Select icon click the object of interest select several objects click the Select icon press and hold the Ctrl key while you click the objects move the selected click on the objects hold down the left mouse button and objectls drag the objectis to the new position resize the selected click one of the object border anchors either in the corners objects or in the middle of a border and drag the box to the new size Note Some Text objects cannot be resized xuri Cell Expansion System W25 User Manual 29 0646 22 AA 107 3
72. The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Alignment toolbar icon functions Objects can be placed in exact positions and sized in relation to other objects The table below describes the function of the Alignment toolbar icons in the Report Editor Toolbar icon Function Align left Matches the left alignment of all selected objects to that of the highlighted object Align right Matches the right alignment of all selected objects to that of the highlighted object Align top Matches the top alignment of all selected objects to that of the highlighted object Align bottom Matches the bottom alignment of all selected objects to that of the highlighted object Adjust to margins Stretches the selected object s to the left and right mar gins Adjust to left margin Adjusts the selected object s to the left margin Adjust to right margin Adjusts the selected object s to the right margin Adjust to center Adjusts the selected object s to the center of the page Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Toolbar icon Function Make same size Adjusts the selected objects to the same size as the high lighted reference object Make same width Adjusts the selected objects to the same width as the highlighted reference object Make sa
73. User Manual 29 0646 22 AA 219 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Error symptom DO control using O seems to react too slow or too fast undershoot ing overshooting Possible cause The bag size setting is incorrect which will op timize the control for another gas volume The control is using manually entered con trol parameters not op timal for the actual case Corrective action Set the bag size to the correct value Use the manual instruction DO control ad vanced DO control advanced O2 to set PID pa rameter mode and Cycle time mode to Auto See Manual instructions on page 47 The automatically se lected control parame ters are not optimal for the actual case Use the manual instruction DO control ad vanced DO control advanced O2 Set PID param eter mode and Cycle time mode to Manual and enter desired vaules for PID parameters and cycle time See Manual instructions on page 47 To get an initial suggestion of the PID parameters look at the automatically selected parameters by selecting them as run data under Tools Customize A gas bubble might be present on the DO sen SOT Tap the DO sensor from the underside to remove the bubble This may require forceful manipulation and the Cellbag bioreactor may need to be partly disconnected from the tray Note The gas bubble may be very difficult to see DO control using O is not using O concentra tion
74. User Manual 29 0646 22 AA 127 4 System control description 4 3 DO control 4 4 DOcontrol In this section xuri Cell Expansion System W25 has automated of DO control using O gt In general PID parameters and cycle times do not need to be entered or tuned Refer to Section 8 8 9 DO measurement on page 276 and Section 8 8 10 DO control on page 279 for information about DO control settings This section contains the following subsections Section See page 4 3 1 O2 control mode 129 4 3 2 Speed control mode 131 4 3 3 DO control transition delays 132 128 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 3 DO control 4 3 1 O2 control mode 4 3 1 O2 control mode Introduction At every execution of the O control a PID regulator computes the O setpoint for the next control cycle There are two different O setpoint ranges If compressed air is selected as gas source to the CBCU the range is 21 to 50 if N is selected the range is 0 to 50 PID parameters and cycle time The control dynamics differ between the different bag sizes and are dependent of the rate of the gas flow The PID parameters and the cycle time are therefore automatically computed with respect to the bag size and gas flow set point To optimize the control behavior the automatic PID parameters are not the same when approaching a new setpoint as when keeping the setpoint The automatic D parameter is always
75. Xuri Cell Expansion System W25 User Manual Page intentionally left blank Table of Contents Table of Contents Hn o ioo a e y een tee Monee E Ene netta Maer omer E Maer EEA etme rr vec E AET 7 IL imponan AIS yr ol TOU paces cehca E O 8 1 2 Mpornant CONCEDIS sissie NEE ei E AE 10 o HS OC amea eee a E ates eee eels 11 2 SYSTEM description csccssccssccsscsrssscsssccsecessseccsecesseessesesesesessesaeecsessessscesses 12 Pr A EOE ON aiena e AE N EE O E A A 13 2 2 Xuri Cell Expansion System W25 rocker nsssssssssssssssssssssssssssessssssrssrsesresrneerierrierrerrerennrrnnenssnnsnsns 16 25 Xur cel Expansion W 25 CBCU seisseen EE e EEEE 26 24 Xuri Cell Expansion W25 PUMI scccissccteccincetsetvcvenaredecccaccusncosceaases sexed e Eai 29 AS LAITE A O eea a AE A EE A OSSEE N 52 2 6 UNICORN software OV ST VOW ssccce ys aesccstcacsosceessessscntesccsccesvenpectcseed did lemstruessesoosteeiaiceUteccdcccoss 36 2 6 1 cennera UNICORN Ko oe gets 6 noesa n aE 37 2 6 2 The UNICORN help u cecsssssssessssessssecssessssssssessssecsssecssscsssessssssssscsssscssuessuecsssssssessssecsssecsnecsseecasecesseces 38 3 The UNICORN software sssrssiicmnenisuiir a 40 2L AMIM TOON aes E E ence ate E 41 ae a V E TO r E ve A A nee en 42 33 Methoas MUNICORN epeen ee a EEE 49 55 1 Menoa edilOr cess cessor cannerresranduereeennieieeta tonsa A E E 50 332 Method CreQtiON icsssiscsesessesnacssvsnsssanshonisann eievsese
76. Y Yes No or Not applicable Transition delay output data Description View from od ae 1 D EEEE D Auto O2 to speed transition The O to speed transition delay time used if delay 02 speed transition delay mode is set to Auto Auto speed to O2 transition The speed to O transition delay time used if delay O02 speed transition delay mode is set to Auto Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 283 8 Reference information 8 8 Control settings 8 8 11 Media control 8 8 11 Media control Media control settings Setting Description The following table shows general settings used for media control Set from PP MI View from PP RD D Media control mode If Media addition is selected the feed pump will run with the flow given by parameter Flow rate until the weight setpoint is reached If Perfusion is selected the feed or harvest waste pump will run with the flow given by parameter Flow rate until the weight setpoint is reached Then both feed and harvest waste pumps will run together keeping the weight setpoint Settings Me dia control Media control general k Weight set point The weight target value Settings Me dia control Media control general Weight devia
77. a CO concentration in air which deviates from the expected value 0 0 It takes up to several minutes for the CO control to reach the new setpoint Minor deviations can appear for example due to temperature variations This is normal Wait 5 to 10 minutes If the deviation is large the offset CO concentra tion can be adjusted Contact GE Healthcare ser vice personnel CO reading keeps drifting down CO gas supply pres Sure is too low Check that the CO supply pressure is between 1 0 and 1 5 bar Erratic CO control CO supply is not prop erly connected Connections for CO gt O N or air have been mixed up Check that the CO gas is properly connected and at the correct pressure 1 0 to 1 5 bar Incorrect setpoint Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Check the setpoint 213 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU O mix Error symptom No O reading Possible cause O sensor is broken Corrective action Contact GE Healthcare service personnel Display shows an O concentration in air which deviates from the expected value 21 0 Minor deviations can appear for example due to temperature variations If the deviation is large the O offset can be adjust ed contact GE Healthcare service personnel O reading keeps drift ing down O gas supply pressure is too low Check that the O su
78. able below Indication White The function is inactive Gray The function is disabled due to higher level control Green The function is active Orange The function needs attention Click on the button to open the related settings and to see more information xuri Cell Expansion System W25 User Manual 29 0646 22 AA 193 5 Operation 5 5 Perform cultivation 5 5 2 Monitor and control the cultivation Indication The function is not working properly Click on the button to open the related settings and to see an explanation of what the problem is Sample the Cellbag bioreactor Samples may be taken several times during the run using the same sampling connector Before taking a sample make sure that the sampling connector is firmly attached to the luer lock coupling of the cellbag bioreactor by screwing it clock wise Follow the instruction below to take samples from the Cellbag bioreactor Step Action 1 Stop the rocking motion from the Process Picture 2 Click the right hand side of the Sampling button to prepare for sampling Result The system will enter sampling mode Note The system will be in sampling mode as many minutes as set in Settings Rock ing Sampling Pause and at an angle set in Settings Rocking Sampling Stop angle in the Process Picture Note For small culture volumes it may be necessary to position the tray into tilt position during sampling Remove the cap from the blue sa
79. according to the information and make sure that the control turns active again Rocker is in sampling mode None If the pH control is using acid or base it is inactive while the rocker is in sampling mode Auto calibration of feed and harvest waste pumps is active Possible cause None If the pH control is using acid or base it is inactive during auto calibration Corrective action No pH reading Off is displayed pH reading is not start ed Open Settings pH from the Process Picture and set Reading to On Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 217 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Error symptom DH reading is inactive 0 0 is displayed and the frame around the Possible cause Repeated errors from the the sensor has turned the reading into Corrective action Read the warning messages for more information Check the possible causes and corrective actions below Process Picture button an inactive state New is red reading attempts are made with asked read ing frequency until a valid reading is re turned or the reading is stopped pH sensor cable isnot Check both ends of the cable Make sure that all connected properly or four pins of the sensor port is gripping the sensor is defective adapter and that the fiber cable is properly con nected to the PH port on the CBCU front panel If both pH and
80. ailable from the Instruction box Repeat this for all red instructions before saving the method Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 7 Troubleshooting 7 6 Software problems 7 6 1 Troubleshooting Method editor Export of a method to a network drive fails The table below describes how to solve a method export problem Problem description Solution Export of a method to a network drive fails Ensure that the destination network drive is mapped and that you have the appropriate access rights Method cannot be created after new Instrument Configuration installation The table below describes how to solve a predefined method creation problem Problem description Solution A new instrument configuration is installed After this The Method Editor must be restarted after im it is still impossible to create a new method porting the new instrument configuration Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 231 7 Troubleshooting 7 6 Software problems 7 6 2 UNICORN System Control 7 6 2 UNICORN System Control User Access Problem description Solution Username and password not ac The UNICORN administrator should check if the user account cepted is locked for example after too many unsuccessful log on attempts The UNICORN administrator can try to set a new password If a password reset does not work the user profile may have to be deleted and a new profile created The log
81. and select Run Data Groups Detailed or e right click and select Customize to customize the appearance of the Run Data pane 48 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 5 Methods in UNICORN In this section This section describes how to create methods and work with predefined methods and text instructions For information about dialogs in the Method Editor that are not de scribed in this manual refer to the online help This section contains the following subsections Section See page 3 3 1 Method editor 50 3 3 2 Method creation 53 3 3 3 Work with methods 5 3 3 4 Text instructions 64 3 3 5 Save a method 72 3 3 6 Scouting 76 3 3 7 Method queues 82 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 49 3 The UNICORN software 3 3 Methods in UNICORN 3 3 1 Method editor 3 3 1 Method editor Introduction In the UNICORN software the instructions to control a bioreactor run can be defined in a method The Method Editor module is used to create or edit such methods User defined methods and phases A method is built up by a number of phases and each phase is built up by a number of instructions Refer to the UNICORN Online Help for more information about methods and phases and how to create a method 50 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 1 Method editor Meth
82. anges to be applied xuri Cell Expansion System W25 User Manual 29 0646 22 AA 233 7 Troubleshooting 7 6 Software problems 7 6 2 UNICORN System Control Problem description Solution You receive the error message Can Check that the rocker power button shows a steady green not connect to system in a network light installation Check that the computer from which you try to establish a connection is logged on to the network Check that the limit of five simultaneous connections to the system has not been exceeded Check the firewall settings on the client computer Refer to UNICORN Administration and Technical manual You receive the error message This error message is displayed if a system is defined and active Warning system occupied when in two different UNICORN database instances and is already trying to connect connected in the other instance It is not recommended to have a system defined and active in more than one UNICORN database instance 234 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 7 Troubleshooting 7 6 Software problems 7 6 3 Troubleshooting Evaluation 7 6 3 Troubleshooting Evaluation Maximum number of curves exceeded The table below describes a problem and its solution Problem description Solution The maximum number of curves 100 are exceeded Delete the unnecessary curves before import when importing curves ing more curves Export of archived result
83. ar view of Xuri Cell Expansion W25 CBCU The illustration below shows the rear panel of a fully configured CBCU xuri Cell Expansion System W25 User Manual 29 0646 22 AA 27 2 System description 2 3 Xuri Cell Expansion W25 CBCU Component UniNet 9 port Description Power connection to the rocker CAN indicator LED Indicates system connection status CAN ID switch Switch for setting the CBCU unit number for system recognition O2 IN Inlet connection for O supply CO2 IN Inlet connection for CO supply AIR N2 Inlet connection for air or Nz supply CAN ID The CAN ID is a unit number used by UNICORN to recognize that the CBCU is connected to the system The CAN ID is set by turning a switch on the CBCU rear panel see illustration above Two possible CAN ID positions are available on the switch marked 1 and 2 respectively The CAN ID should always be set to position 1 28 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 4 Xuri Cell Expansion W25 Pump 2 4 Xuri Cell Expansion W25 Pump Introduction xuri Cell Expansion W25 Pump is a peristaltic pump unit that includes two roller pumps It pumps fluid for feed harvest waste and pH control with acid and base The dimensions and weight of the pump are listed in Section 8 2 Component specifications on page 238 Front view of the pump The illustration below shows the front panel of th
84. ase can be entered using the manual instruc tion pH control advanced acid base This will override the automatically computed flow rates xuri Cell Expansion System W25 User Manual 29 0646 22 AA 125 4 System control description 4 2 pH control 4 2 2 Acid Base control mode Dead band 126 A dead band zone is defined for acid and base individually When the control output is in the dead band zone the pumps will not run Control Acid Base output dead dead band band Acid Base The dead band is removing all acid base injections with a pump run time less or equal to a given percentage of the cycle time The default dead band is set to 2 0 which means that only injections with a pump run time greater than 2 0 of the cycle time will be executed The dead band prevents the control to toggle between small acid and base injections Using dead band values close to zero will give a precise control but also a risk of unnec essary toggling between acid and base The greater the dead band values are the less precise the control will be Using a large dead band might even lead to that the setpoint is never reached The default value of 2 0 has been proven to be a good compromise The dead band is set using the manual instruction pH control advanced acid base Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 2 pH control 4 2 3 pH control transition delays 4 2 3 pH control trans
85. ation in the gas mix and by chang ing the rocking speed e The software switches automatically between O2 and Speed mode start ing in 02 mode The control cycle The measurements and control actions are synced When starting the pH or DO control the measurement cycle time is set by the control and overrides the entered reading cycle time After every new measurement the control is executed When the pH control executes it sets the CO setpoint for the next control cycle CO2 mode or starts the acid or base pump Acid Base mode for a limited time 0 75 of the next control cycle New measurement Ne ontrol execution Control cycle When the DO control executes it sets the O setpoint for the next control cycle 02 mode or sets the rocking speed setpoint for the next control cycle Speed mode Automatic and manual mode The pH and DO control software automatically calculates the following parameters suitable for the actual control case taking into account the bag size and gas flow setpoint e Cycle time e PID parameters e Acid and base flow for pH control e Transition delay time between CO and base in the CO2 Base control scheme and O and speed in the 02 Speed control scheme xuri Cell Expansion System W25 User Manual 29 0646 22 AA 119 4 System control description 4 1 pH and DO measurement and control 4 1 3 pH and DO control principles Note In DO control Speed mode there are only ma
86. bing are needed as when calibrating manually and measuring the pumped volume in a measuring glass The auto calibration assumes a density of the liquid equal to 1 So when using auto calibration with a media of other density the flow rate given in L day shall be seen as given in kg day instead Note During auto calibration it is of high importance not to touch the system and that the bench the rocker is placed on is unaffected by the rocking as this can disturb the weight measurements To get an indication of the auto calibration result inspect the feed and harvest calibration factor curves in the curve data chart If running perfusion at the weight setpoint with both pumps running and the auto cali bration disabled an auto calibration can be immediately triggered by enabling autocal ibration If an auto calibration was disturbed and needs to be redone the auto calibration can be disabled and then enabled again to trigger a spontaneous auto calibration Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 4 Media control 4 4 2 Perfusion Auto calibration cycle The figure below shows auto calibrations during a perfusion run The first auto calibration is done when the weight setpoint is reached This calibration replaces the manual cali bration needed before starting if auto calibration is disabled After that the auto calibra tion is repeated with the interval given in the manual instruction M
87. blems 7 6 2 UNICORN System Control Solution Either stop the manual run which will allow the method run to start or start another method This will add the second method to the list of running manual run The start protocol cannot be complet methods and the first method is allowed to start ed and the method run cannot start at this point This is because the alarm must be acknowledged first but no message about this is issued System connections Solution Problem description The connections are not available i e e the selection checkbox is grayed out Check if the system has been deactivated e Check that the power to the system is turned on e Check that the rocker power button shows a steady green light e Check the connection between the client computer and the system e Check the firewall settings on the client computer Refer to UNICORN Administration and Technical manual The connections are not available 1 even though Switch off the system 2 Exit UNICORN e the connection between the PC and system appears to be correct 3 and Restart the system 4 Log on to UNICORN e the power is turned on A system is not available when you E attempt to establish a connection Check that you have access rights to the system Access rights are not automatically assigned for a newly defined system e The system may not be active e Log off and log on again for access rights ch
88. calibration will start Media additon is select ed as control mode Auto calibration is only done in perfusion mode The second auto cali bration does not seem to occur After auto calibration the flow of one or both of the pumps is no longer correct The Auto calibration cycle is set to a long cycle time that has not yet elapsed The weight measure ments done by the auto calibration routine might have been dis turbed by something for example streched tubing or someone touching the system Check what the actual Auto calibration cycle is by selecting it as run data under Tools Customize Use the manual instruction Media control Media control general and set Auto calibration cycle to desired value Also select Auto calibration cycle timer as run data under Tools Customize to see how many hours that are left to the next auto calibration Recalibrate the pump pumps by first disabling then enabling the auto calibration in the Process Picture A new auto calibration will then start During the auto calibration be sure that nothing interfers with the system 224 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Error symptom The media control is automatically turned off Possible cause None of the pumps started by the media control are no longer running xuri Cell Expansion System W25 User Manual 29 0646 22 AA 7 Troubleshooting 7 3 Xuri Cell Expansion W25
89. ce Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 4 Media control 44 Media control In this section The media control has two modes Media addition and Perfusion The two modes are described in the following sections A complete description of all settings and output data is found in Section 8 8 11 Media control on page 284 This section contains the following subsections Section See page 4 4 1 Media addition 134 4 4 2 Perfusion 135 4 4 3 Deviation alarm 139 4 4 4 Media control inactivation 140 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 133 4 System control description 4 4 Media control 4 4 1 Media addition 4 4 1 Media addition Media addition principles The media addition mode will fill the bag by running the feed pump at the desired feed rate until the weight setpoint is reached To keep the desired feed rate the media control uses a transitional weight setpoint that is ramped towards the given weight setpoint It is called momentary weight setpoint and the ramping speed corresponds to the feed rate The feed flow is adjusted by a regulator to keep the weight measurement at the momen tary weight setpoint Weight setpoint Momentary weight setpoint Weight measurement Weight Start Time When the weight setpoint is reached the feed pump is stopped but the control is still on and will start the feed pump again if the we
90. col 1 Type the Result name and change the Location of where the result is saved if necessary 2 Select the correct Bag size 3 Click OK Result The manual run starts Start a method run The instruction below describes how to start a method run Tip Step It is possible to have a method run going for example a method based on the predefined method WAVE 25 template and at the same time control the system from the Process Picture Action Open the System Control module and click the icon Open Method Navigator Method Navigator Open wa ie Methods Method Qi Folder name System s Eric Methods E Method Systema FL Method Systema Results Select the method to run and click the Run icon Pd Result The Start Protocol dialog for the method run opens On the displayed page in the Start Protocol add requested input and make appropriate changes If necessary Click Start Result The method run starts Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 179 5 Operation 5 4 Prepare cultivation 5 4 1 Start a run Hold or stop the run To interrupt a method during a run you may use the Hold or End icons in System Control A held method run can be resumed by using the Continue icon See the instructions in the table below At the end of a method the run stops automatically All functions stop including rocking and data logging and an acoustic end
91. con A Mag Result The Generate Report dialog box opens 5 e Click the New button Result The Customize Report window opens in Edit mode 100 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format The Edit mode window The illustration below shows the Customize Report window in Edit mode with a blank report open g AddPage Cowen Ext Dk TSHA GALAT hee aia 4 Page 1 of 1 Ed Mode Toolbar commands in the Customize Report window The table below describes the different functions of the toolbar command buttons in the Customize Report window Toolbar button Function Preview Edit mode This button toggles between a print preview of the report and the Edit mode Prev Page This button displays the previous page or pair of pages if there is more than one page Next Page This button displays the next page or pair of pages if there is more than one page One Page Two Pages This button toggles between single page view and pairs of pages view if there is more than one page xuri Cell Expansion System W25 User Manual 29 0646 22 AA 101 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Toolbar button Function Select the magnification of the view in this droplist Add Page This button adds a blank page to the report Delete Page This button deletes the current
92. connector The full configuration mixes air N gt O and CO gas and contains O and CO sensors a mass flow controller an optical pH sensor reader and an optical DO sensor reader Three configurations are available e CO gt O and pH e CO O and DO e CO O gt pH and DO full configuration The dimensions and weight of the CBCU are listed in Section 8 2 Component specifications on page 238 Front view of Xuri Cell Expansion W25 CBCU The illustration below shows the front panel of a fully configured CBCU The configuration of your CBCU may vary from the configuration shown below Xuri Cell Expansion W25 CBCU pH GAS MIX OUT DO i R o a a 1 2 3 4 26 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 3 Xuri Cell Expansion W25 CBCU Component Description pH port Connector for pH sensor fiber cable GAS MIX OUT Gas outlet for connection to Cellbag bioreactor Status LED Indicates the CBCU operating status DO port Connector for DO sensor fiber cable Status LED The status LED indicates the CBCU operating status according to the following table Light indicator Description Steady green light The CBCU is ready for operation Green blinking light The CBCU is operating Red blinking light Indicates an internal error but the CBCU is still operating Steady red light Indicates an internal error and the CBCU is not operating Re
93. ctions on page 47 Check what the actual deadband values are by selecting them as run data under Tools Customize Use the manual instruction pH control ad vanced pH control advanced acid base De crease the deadband value for acid and or base See Manual instructions on page 47 216 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Error symptom In control scheme CO2 Base the transi tions between CO and base are longer or shorter than desired Possible cause The automatically computed transition delay time is not opti mal for the actual cell culture 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Corrective action Check what the actual trasition delay time values are by selecting them as run data under Tools Customize Use the manual instruction pH control advanced pH control transition delay Set CO2 Base transition delay mode to Manual and enter desired values for transition delays See Manual instructions on page 47 The control is using a manually entered tran sition delay time not optimal for the actual case Use the manual instruction pH control ad vanced pH control transition delay Set CO2 Base transition delay mode to Auto See Manual instructions on page 47 pH control is inactive pH reading Error symptom Some of the criteria for running the pH control are no longer fulfilled Read the message in the pop up dialog If possible take action
94. d DO setpoint lower dead band there will be no control actions Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Transition delay settings The following table shows the transition delay settings used for DO control Setting 02 Speed transition de lay mode Description When Auto is selected the system will automatically set the transition delay time The automatically comput ed transition delay times can be dis played as run data When Manual is selected the system will use the manual transition delay times set by this instruction Set from MI DO control transition delays View from PP RD CD A hand symbol is shown if set to Manual Manual O2 to speed delay The transition delay time from O to speed control used if O2 Speed transition delay mode is set to Man ual DO control transition delays Manual speed to 02 delay 282 The transition delay time from speed to O control used if O2 Speed tran sition delay mode is set to Manual DO control transition delays Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 10 DO control Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data
95. d cells Instructions Follow the instructions below to fill the Cellbag bioreactor with culture medium Step Action 1 Reduce the gas flow to avoid high pressure alarm Note Continue gas flow to the headspace to keep the Cellbag bioreactor inflated xuri Cell Expansion System W25 User Manual 29 0646 22 AA 185 5 Operation 5 4 Prepare cultivation 5 4 4 Addition of culture medium Step Action 2 Open Settings Weight from the Process Picture in System Control Front Left 25 0 Front Right 75 0 36 Rear Left 25 0 Rear Right 25 0 Net Weight 0 00 kg Tare 3 Check that the weight distribution is even by reading the weight percentage values for the rocker feet The values should not differ more than 5 and the optimal weight distribution is 25 on each load cell Turn the adjustable foot if necessary see Adjustable foot on page 18 Note The tray must be in horizontal position 4 Make sure that the lid and all other equipment that will be used during the run is placed on the tray and that no tubing weighs down the tray Click Tare 5 Open Rocking from the Process Picture Change the Stop Angle to 12 0 and click OK Note The risk that air bubbles are trapped by the optical sensors is minimized with the Cellbag at an angle The angle can be adjusted to make sure that the medium reaches all optical sensors during filling 6 Slowly transfer the desired volume of medium into the C
96. d dialog opens Find what Stop acid pumpi Find Nest Direction Cancel C Match whole word only C Up 5hit F3 C Match case Down F3 96 xuri Cell Expansion System W25 User Manual 29 0646 22 AA Step 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 3 Run documentation Action 3 e Type the text you want to locate in the Find what textbox Note Your previous search text may be shown in this box if you have used the search function before e Select additional search criteria Match whole word only Match case Search Up Search Down e Click the Find Next button Result The first log entry where the search text is found is marked Save the method used for the run as a new method You can save the method with the variables that were used for the run as anew method as described below Step Action 1 2 Select the Text Instructions tab e Click the Save as button Result The Save As dialog box opens e Select the appropriate destination folder e Type aname in the Name text box e Select a system from the System droplist Result The method is saved xuri Cell Expansion System W25 User Manual 29 0646 22 AA 97 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 4 Generate and print a predefined report format 3 4 4 Generate and print a predefined report format Introduction This section describes how to generate and print a report using a f
97. degrees This position can also be set by executing the manual instruction Rocker Prepare for tilt See illustration below In order to facilitate sampling and harvest it is possible to position the tray with the at tached Cellbag bioreactor into an upright position called tilt position For visibility purposes the tray is shown without the Cellbag bioreactor attached in the images below Step Action 1 Select the largest possible angle in UNICORN i e 12 degrees or prepare for tilt as described above xuri Cell Expansion System W25 User Manual 29 0646 22 AA 23 2 System description 2 2 Xuri Cell Expansion System W25 rocker Step Action 2 Grab the textured grip area on each side of the tray and pull the tray towards yOu The illustration below shows the tilt position 24 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 2 Xuri Cell Expansion System W25 rocker Illustration of filter heater The filter heater prevents occurrence of condensation and clogging of the outlet gas filter of the Cellbag bioreactor Description Filter heater Connector for connection to the rocker Filter heater stand Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 25 2 System description 2 3 Xuri Cell Expansion W25 CBCU 2 5 Xuri Cell Expansion W25 CBCU Introduction The control unit Xuri Cell Expansion W25 CBCU is connected to the rocker via a UniNet 9
98. dition of culture medium 185 5 4 5 Equilibrate the culture medium 188 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 4 Prepare cultivation 34 1 Starra run 5 4 1 Start a run Introduction This section describes how to start a manual run or to start a run using a previously defined method The data collection begins when a run starts For further information on method creation refer to Section 3 3 Methods in UNICORN on page 49 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 177 5 Operation 5 4 Prepare cultivation 5 4 1 Storta run Start a manual run The instruction below describes how to start a manual run Step Action 1 Acknowledge a change of any settings for example 1 Click the Cellbag icon E i ia we Poe ig Se Cellbag 10L S 2 Enter pH and DO calibration data Bag size Cellbag 10L Bag sensor calibration data Cellbag pH Cmin 58 00 Left Single Cmax 30 00 Right ae a Weight CdpH 1 00 Rocking CTemp 20 DO n me Clhp 26 00 Clht 37 00 Cizt 37 00 Calp 1000 i Cancel 3 Click OK Result The Start Protocol dialog for the manual run opens Result name Manual Run 010 Location DefaultHome Bag details Bag size Cellbag 20L 178 xuri Cell Expansion System W25 User Manual 29 0646 22 AA Step 5 Operation 5 4 Prepare cultivation 34 1 Stant arun Action On the displayed page in the Start Proto
99. ditions will therefore be obtained at high altitudes as compared to at sea level 21 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 1 pH and DO measurement and control 4 1 2 pH and DO reading 4 1 2 pH and DO reading Introduction UNICORN is used to start the pH and DO reading and to set the time between measure ments Start the reading To start the pH reading set Reading On under Settings pH in the Process Picture or use the manual instruction pH sensor Start pH reading To start the DO reading set Reading On under Settings DO in the Process Picture or use the manual instruction DO sensor Start DO reading Note Pressing the right hand side of the pH or DO button in the Process Picture will start both the reading and the control pH and DO reading cycle time The reading cycle time is the time between two consecutive measurements Reading New measurement cycle The reading Cycle time is set from the Process Picture under Settings pH and Set tings DO or with the manual instructions Set pH reading cycle time or Set DO reading cycle time When the control is off the reading cycle time setting defines time between measure ments When the control is on the reading cycle time will be overridden by the control cycle time Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 117 4 System control description 4 1 pH and DO measurement and control 4 1 3
100. duration mode before it automatically starts to rock again See help text for Enter sampling mode instruction to learn more about sampling mode Cellbag size The Cellbag size defined in total Settings Cellbag size Y total volume volume liquid and head space Cellbag Digital Sets the outputs of the digital Digital output Y output output ports Rocking Defines the share of the rocking Set rocking Y motion cycle where the speed curve is motion sinusoid The lowest value 15 provides a rocking motion simi lar to the Wave 20 50 system The highest value 100 will give a completely sinusoidal speed curve Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 249 8 Reference information 8 8 Control settings 8 8 1 Rocking control Setting Description Set from View from PP RD Auto start By setting this parameter to Not System set mode resume activity rocking or tings Auto heating will not start automati start Rocker cally at startup of the system at any time When the parameter is set to Resume activity rock ing and heating will start auto matically at startup if it was on when the system was shut down without given an END command Prepare for If No the rocking platform will System set tilt at END not prepare for tilt at system tings Rocker END if Yes the rocking platform will prepare for tilt at system END Note In the table the
101. e 188 When required CO gt O gt scale and temperature are calibrated at manufacturing It is recommended to calibrate these parameters at least annually Calibration of CO and O is complex and requires expert knowledge Calibration Interval When required Contact GE Healthcare service personnel for assistance if needed Note It is recommended to perform scale calibration after moving the rocker or if working with a load that differs considerably from the load at the last calibration For Calibrate High point use a weight that is as close as possible to the load to be applied to the tray during use Temperature When required Contact GE Healthcare service personnel for assistance if needed CO sensor CO calibration require special competence and may impair system performance if performed inappropriate Contact GE Healthcare service personnel for assistance O sensor O calibration require special competence and may impair system performance if performed inappropriate Contact GE Healthcare service personnel for assistance xuri Cell Expansion System W25 User Manual 29 0646 22 AA 201 6 Maintenance 6 1 Calibrations Calibration instruction Follow the instructions below to perform a calibration The example is a scale calibration Step Action 1 2 2 4 e Select System Calibrate in System Control or e click Calibrate in the Process Picture Settings menu for the monito
102. e Scouting parameters table and click LlsetBun_ Result A new row is added below the selected run The variable value from the selected row is copied to the new run Each chosen variable is displayed in a separate column Method Settings METHOD SETTINGS Run Included Rocking Speed APH 1 20 0 2 20 0 e Inthis example click in the Rocking Speed RPM column for the appro priate run and edit the rocking speed value Note Changing variable values in the scouting scheme does not change the values in the Variable List in the Duration and Variables dialog accessed from the Method Editor or in the text instructions The actual variable values used for each run in the scouting scheme are saved in the result file To change the default values the variable values must be edited in the Phase Properties pane e Repeat until all runs are included using the correct variable values Note The scouting scheme can also be edited just prior to starting the method run in the Start Protocol Here variable values can be changed and indi vidual runs included or excluded Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 79 3 The UNICORN software 3 3 Methods in UNICORN 5 5 6 Scouting Step Action 5 To insert a series of runs e Click in the appropriate variable column in the Scouting parameters table and click L_Set _ This button is activated for variables with continuous values such as flow rates or pressure limits R
103. e l parameter used if PID parameter mode is set to Auto Auto DO 02 D The D parameter used if PID parameter mode is set to Auto Auto DO O2 Cycle time The cycle time used if Cycle time mode is set to Auto Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable Speed control mode settings The following table shows Speed control mode settings used for DO control Setting Description Set from View from MI d na gt De D Max speed The maximum rocking speed setpoint DO control Y allowed for the DO control speed Min speed The minimum rocking speed setpointal DO control Y lowed for the DO control speed Speed step The change of rocking speed setpointat DO control Y each control action when DO reading is speed outside dead band xuri Cell Expansion System W25 User Manual 29 0646 22 AA 281 8 Reference information 8 8 Control settings 8 8 10 DO control Setting Description Set from View from MI PP RD CD DO control speed The cycle time of the DO speed control i e the time between each control ac tion Control cycle DO control speed Upper dead When DO reading is between DO setpoint band and DO setpoint upper dead band there will be no control actions DO control speed Lower dead When DO reading is between DO setpoint band an
104. e layout is changed in the Page Setup dialog The table below describes how to set up the page layout Step Action l e Double click anywhere on the report page not on an object or e Right click not on an object and choose Properties from the shortcut menu or e Choose the Edit Page setup menu command Result The Page Setup dialog box opens Page Setup Fage Setup Footer I First Header Header Margins Top Units Bottom Lett Right Ci cm inch Settings C Same header on all pages we a frame around the pages 2 e Type new values for the Margins if necessary e Select the appropriate Settings and Unit Note An extra Header tab will appear if you de select the option to have the same header on all pages The First Header tab is used for the first page header only and the Header tab is used for all subsequent pages 104 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Step Action 2 e Click the First Header tab e Select all the items you want to include in the header from the Select Items list e Click the Font button to change the font for all items if necessary 4 e Type header text in the Free text box and click the Font button to alter the default font if necessary This text will be placed on top of the header e Type the report title in the Rep
105. e light is blinking green while shutting down Note If the rocker fails to shut down keep the power button pressed in more than 4 seconds to force a shutdown 3 Clean the bioreactor system units Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 199 6 Maintenance 6 Maintenance In this chapter 200 This chapter describes the required maintenance procedures for Xuri Cell Expansion System W25 It also gives an overview of the calibration procedures needed for the system to function properly Maintenance manager The maintenance manager in UNICORN keeps track of the usage of different components and shows alerts when it is time for maintenance and service For detailed information about the maintenance manager refer to UNICORN Administration and Technical manual Section See page 6 1 Calibrations 201 6 2 Pump calibration 203 6 3 Cleaning 205 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 6 Maintenance 6 1 Calibrations 6 1 Calibrations Calibration schedule For the system to function properly several calibrations may be performed See tables below Before each cultivation Calibration Interval Pump Before cultivation see Section 5 4 2 Pre culture preparation on page 182 DO sensor Before cultivation see Section 5 4 5 Equilibrate the culture medium on page 188 pH sensor Before and during cultivation if required see Section 5 4 5 Equilibrate the culture medium on pag
106. e pump Xuri Cell Expansion W25 Pump Description Pumphead flip top Pumphead Status LEDs for pumping function per pumphead Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 29 2 System description 2 4 Xuri Cell Expansion W25 Pump Status LEDs The status LEDs indicate the pumping function status according to the following table Light indicator Description Steady green light The pumping function is ready for operation Green blinking light Pumping is ongoing Red blinking light Indicates an internal error but the pump is still operating Steady red light Indicates an internal error and the pump is not operating properly Rear view of the pump The illustration below shows the rear panel of the pump 1 2 3 29 0645 71 Xuri Cell Expansion W25 Pump Code no 29083569 Voltage 32 V DC UniNet 9 Serial no XXXXXXX Mfg Year 2013 AZ BEL Max Power 32 W AZOO _ GE Healthcare Bio Sciences AB Coo suomi Vy Component Description UniNet 9 port Power connection to the rocker CAN indicator LED Indicates system connection status CAN ID switch Shows the unit number of the pump for recognition by the system 30 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 4 Xuri Cell Expansion W25 Pump CAN ID The CAN ID is a unit number used by UNICORN to recognize the particular pump unit that is connected
107. e result as Click Execute Auto update of parameters during run 130 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 3 DO control 4 3 2 Speed control mode 4 3 2 Speed control mode Introduction At every execution of the speed control the DO reading is compared with the DO setpoint If the DO reading is below the DO setpoint the rocking speed setpoint is increased If the DO reading is above the DO setpoint the rocking speed setpoint is decreased The settings of the DO speed mode can be set from the Process Picture or by using the manual instruction DO control speed Speed step The Speed step parameter defines the size of the change of the rocking speed setpoint Maximum and minimum rocking speed The Max and Min speed parameters define the interval in which the DO controller can set the rocking speed setpoint Dead band The upper and lower dead band parameters defines the dead band interval around the DO setpoint When the DO reading is in that interval no change of the rocking speed setpoint will take place Cycle time The cycle time parameter sets the cycle time of the DO speed control This cycle time will override the DO reading cycle time when the DO control is on Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 131 4 System control description 4 3 DO control 4 3 3 DO control transition delays 4 3 3 DO control transition
108. e system functions This chapter contains the following sections Section See page 4 1 pH and DO measurement and control 114 4 2 pH control 121 4 3 DO control 128 4 4 Media control 133 4 5 Recommended operating conditions 141 4 6 System verification 144 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 113 4 System control description 4 1 pH and DO measurement and control 4 1 pHand DO measurement and control In this section This section describes the pH and DO measurement and control principles of Xuri Cell Expansion System W25 This section contains the following subsections Section See page 4 1 1 pH and DO measurement principles 113 4 1 2 pH and DO reading 117 4 1 3 pH and DO control principles 118 114 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 1 pH and DO measurement and control 4 1 1 pH and DO measurement principles 4 1 1 pH and DO measurement principles Introduction In order to monitor and control DO and pH of a culture using Xuri Cell Expansion System W25 it is necessary to use Cellbag bioreactors equipped with optical pH and DO sensors The optical pH and DO sensors comprise a luminophoric dye immobilized on a substrate that is integrated into the Cellbag bioreactor Pulses of light from a LED on the pH mon itor produce a responsive light signal from the sensor that indicates the pH or DO sur rounding the sensor An optical fiber cable transfer
109. eater connectors UniNet 9 ports USB ports Ethernet connector Power connector xuri Cell Expansion System W25 User Manual 29 0646 22 AA 19 2 System description 2 2 Xuri Cell Expansion System W25 rocker Tray and lid sizes Trays and lids are available in the different sizes listed below Illustrations of tray and lid The illustration below shows the rocker with Tray 50 attached 1 2 Description Bag clamp upper Bag clamp opener one in each upper corner Bag clamp opener one in each lower corner Bag clamp lower The illustration below shows the rocker with Tray 50 and Lid 50 mounted 20 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 2 Xuri Cell Expansion System W25 rocker Description Rocker base Lid Tray Tubing exit Hatch Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 21 2 System description 2 2 Xuri Cell Expansion System W25 rocker The illustration below shows the rocker with Tray 10 and Lid 10 mounted 22 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 2 Xuri Cell Expansion System W25 rocker Prepare for tilt Tilt position When the system enters END mode the tray prepares for tilt by default It can be changed in System Settings Rocker Prepare for tilt at END This position is the mechanical end position of the rocker which is 14
110. edia control general When changing the flow rate or the weight setpoint a new auto calibration will be done Thereafter they will continue with the given frequency Weight Weight setpoint A Weight measurement Start Auto calibration cycle E Feed only Auto calibration Auto calibration Harvest only Auto calibration Time Auto calibration criteria The calibration volume is depending on the flow rate and is computed to make an optimal compromise between pump time and accuracy The pump calibration run time is limited between 2 and 30 minutes for each pump A low flow rate will give a long calibration time and a high flow rate will give a short calibration time If the flow rate setpoint is below 3 43 L day no auto calibration can be done since achieving a volume large enough to get the desired accuracy would demand a pump run time longer than 30 minutes If auto calibration is enabled and the criteria above cannot be fulfilled the media control cannot start unless the flow rate is increased or the auto calibration is disabled xuri Cell Expansion System W25 User Manual 29 0646 22 AA 137 4 System control description 4 4 Media control 4 4 2 Perfusion Recommended weight setpoints The media control will not start if the weight setpoint is outside the possible range shown in the table below Cellbag size L Recommended weight setpoint kg Possible weight setpoint kg SW limited 0 5 1 0
111. ellbag bioreactor using a pump or gravity flow Tip To automatically fill the bag to a desired weight use Media Addition from Settings Media control in the Process Picture 7 Increase the gas flow to operation setpoint 186 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Step Action 5 Operation 5 4 Prepare cultivation 5 4 4 Addition of culture medium 8 If pH and or DO control will be used check the pH and DO sensor labels on the Cellbag bioreactor for sensor calibration data Note Do not start pH and or DO reading until the medium is equilibrated The sen sors do not give reliable measurements until then 1 Open Settings Cellbag Left Single 2 Make sure the correct Bag size is selected 3 Enter the Bag sensor calibration data Bag size Cellbag 10L Bag sensor calibration data pH Cmin 58 00 Cmax 30 00 CpHo 8 00 CdpH 1 00 CTemp 20 DO Clhp 26 00 Clht 37 00 Cizp C60 Cizt 37 00 Calp 1000 K Lox Cancel 4 Click OK Cellbag Left Single Right Weight Rocking Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 187 5 Operation 5 4 Prepare cultivation 5 4 5 Equilibrate the culture medium 5 4 5 Equilibrate the culture medium Important Instructions Equilibration of the culture medium for a minimum of 2 hours is important in order to reach the desired operating conditions and for the optical sensors to function properly Standard conditions for 100
112. elow the setpoint the O gt control 02 concentration to the Cellbag is allowed to be without triggering the deviation alarm Deviation A parameter that defines for alarm delay how long the O concentration to the Cellbag can be outside the allowed deviation limits before the deviation alarm is triggered Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable 258 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 5 O mix control O mix control output data Data Description View from RD CD Pp The measured O concentration from the CBCU Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 259 8 Reference information 8 8 Control settings 8 8 6 Pump control 8 8 6 Pump control General pump settings The following table shows general settings used for pump control Setting Description View from ed ey D E D Pump 25 1A Assigns pumphead 1A to Acid System settings Set role Base Feed or Harvest Waste Pump 25 1A role Pump 25 1B Assigns pumphead 1B to Acid System settings Set role Base Feed or Harvest Waste Pump 25 1B role Pump 25 2A Assig
113. ement accuracy 0 6 1 of read value within 0 to 50 O gt when mixed only with air N O control range 0 to 50 O when mixed with No 21 to 50 O when mixed with air Total O accuracy 1 2 1 of read value O pH measurement range DH 4 5 8 5 OH measurement accuracy 0 05 for pH 6 75 7 25 0 1 for pH 6 5 7 5 OH control range pH 6 0 8 0 Total pH accuracy Process value refer 0 1 for pH 6 75 7 25 ence pH 0 15 for pH 6 5 7 5 DO measurement range 0 to 250 air saturation DO measurement accuracy 5 air saturation excluding atmospher ic pressure variations DO control range 0 to 100 air saturation Total DO accuracy Process value refer 10 air saturation excluding atmo ence pH spheric pressure variations Xuri Cell Expansion W25 Pump specifications The table below he table below lists the system specifications of Xuri Cell Expansion W25 Pump Parameter Pump flow rate range 100 ml 144 I day 0 07 100 ml min 240 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Parameter Pump flow accuracy 8 Reference information 8 2 Component specifications 0 1 5 of read value after calibration Accumulated pumped volume accuracy 10 of measured volume Supported tubing dimensions Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Inner diameter 0 5 to 4 8 mm 1 50 to 3 16 Wall thickness 1 6 mm 1 16
114. entire Xuri Cell Expansion System W25 Operating Instructions before installing operating or maintaining the instrument Always keep the Operating Instructions at hand when operating Xuri Cell Expansion System W25 Do not operate Xuri Cell Expansion System W25 in any other way than described in the user documentation Otherwise you may be exposed to hazards that can lead to per sonal injury and you may cause damage to the equipment Intended use xuri Cell Expansion System W25 is intended to be used as laboratory and manufacturing equipment for cell cultivation Prerequisites In order to follow this manual and use the system in the intended manner it is important that e you have a general understanding of how the client computer and Microsoft Win dows operating systems work e you are acquainted with the use of general laboratory equipment and with handling of biological materials e you have read and understood the Safety instructions chapter in the Operating Instructions e the system is installed according to the instructions in the Operating Instructions e auser account has been created according to UNICORN Administration and Technical manual 8 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Safety notices 1 Introduction 1 1 Important user information This user documentation contains WARNINGS CAUTIONS and NOTICES concerning the safe use of the product See definitions below Warnings
115. ering of the tubing Adjust tubing holder position Use a pointed device such as a ball pen to adjust the tubing holder positions on both sides of the pumphead Follow the instructions below to change from outer to inner position Step Action 1 Make sure that the pump is not running 2 Open the flip top of the pumphead completely 162 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 4 Prepare the pump Step Action 3 Place the pointed device in the small depression on one side of the pump head 4 Press down and towards the back of the pumphead 6 Release the pressure and the jaw rises into the new position xuri Cell Expansion System W25 User Manual 29 0646 22 AA 163 5 Operation 5 1 Prepare the system 5 1 4 Prepare the pump Step Action 7 Repeat the above steps to adjust the tubing holder on the other side of the pumphead To change the tubing holder position from inner to outer position carry out the same procedure but press towards the front of the pumphead NOTICE Make sure that the tubing holder position is not caught in between the inner or outer position as this may cause erroneous flow rates and abnormal tube wear Load tubing Follow the instructions below to load tubing in the pumphead Step Action il Make sure that the pump is switched off 2 Open the flip top of the pumphead completely E Ad
116. esult The Insert Series dialog for the selected variable opens Insert Series For Rocking Speed x Enter start value how much the value shall be Increased for each run and how many rung to add Range 2 0 40 0 Start value Shep by Number of runs m 0 0 Set as integer values 6 In the Insert Series dialog e Enter Start value Step by and Number of runs In this example 20 2 and 5 Result The Scouting parameters table is updated Scouting parameters Method Settings METHOD SETTINGS Run Included Rocking Speed RPH 1 20 0 2 20 0 3 23 4 24 0 5 26 0 E 28 0 80 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 5 5 6 Scouting Step Action 7 Alternatively to enter either consecutive or non consecutive integer values e Check the Set as integer values box in the Insert Series dialog Result The following alternative Insert Series dialog for the selected variable opens Insert Series For Rocking Speed x Enter variable ranges separated by commas For example 1 3 5 12 Range 2 0 40 0 e Enter the appropriate range for example 20 23 25 28 Result The Scouting parameters table is updated Scouting parameters Method Settings HMETHOD SETTINGS Run Included Rocking Speed RPH 1 20 0 2 20 0 3 21 0 4 22 0 5 23 0 E 25 0 7 26 0 8 27 0 q 28 0 8 Click OK in the Scouting dialog to save the scouting
117. ext instructions 64 Text instructions Add a new instruction 64 Help 64 Syntax errors in 230 Text editing a method 64 Tilt position 23 Transition delays 127 132 Tray Attach 153 Detach 155 Size 20 Troubleshooting access to UNICORN func tions 233 export of archived re sult 235 maximum number of curves exceeded 235 system or computer connec tions 234 user access 232 Tubing Load tubing 164 Tubing holder position 161 U UNICORN 36 Administration module 41 connect to system 172 Evaluation module 87 Log on 170 Method Editor module 50 Start 170 System Control module 42 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA V Variables Breakpoints or gradient lengths 67 Defining 66 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Index Identification in text instruc tions 67 W Wave motion Illustration 14 291 Page intentionally left blank For local office contact information visit www gelifesciences com contact GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Sweden www gelifesciences com xuri imagination at work GE imagination at work and GE monogram are trademarks of General Electric Company Cellbag UNICORN and Xuri are trademarks of GE Healthcare companies This product or portions thereof is sold under a sub license from Sartorius Stedim Biotech under US 6 673 532 US 7 041 493 and or its foreign equivalents Clave is a
118. following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Rocking control output data Description View from PP RD CD Rocking speed measurement The actual rocking speed measured by the rocker Y Y Y Time left in sampling mode The time left until the rocker starts to rock whenin Y sampling mode Unit s Weight measurement The weight measured by the rocker If tared correct Y Y Y ly it should correspond to the weight of the content of the Cellbag Detected tray type The tray type detected by the system Y Weight share front left Percentage of the weight load on the front leftload Y cell 250 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 1 Rocking control Description View from d ae 1 D EEEE D Weight share front right Percentage of the weight load on the front right load cell Weight share rear left Percentage of the weight load on the rear left load cell Weight share rear right Percentage of the weight load on the rear right load cell Digital input 1 Shows if the digital input 1 is O or 1 Digital input 2 Shows if the digital input 2 is O or 1 Digital input 3 Shows if the digital input 3 is O or 1 Digital input 4 Shows if the digital input 4 is O or 1 Analog input Shows the analog in
119. following from the Process Picture e click Settings DO and set Reading On e click Settings pH and set Reading On Wait until the values are stable 6 Perform the DO calibration 1 Select System Calibrate in System Control 2 Select the DO sensor in the Monitor to calibrate drop down menu 3 Enter percentage of air saturation 90 100 depending on CO con centration in the Enter reference DO field 4 Click Calibrate 5 Close the Calibration dialog 7 Click the right hand side of the Sampling button to prepare for sampling Result The system will enter sampling mode Note The system will be in sampling mode as many minutes as set in Settings Rock ing Sampling Pause and at an angle set in Settings Rocking Sampling Stop angle in the Process Picture 8 Take a sample to verify that the pH value shown by the system matches the pH measured with a calibrated reference instrument If the deviation is larger than approximately 0 5 pH units make sure that no air bubble is present For instructions on how to remove air bubbles see pH reading on page 217 9 If the deviation is less than 0 5 pH units do the following 1 Select System Calibrate in System Control 2 Select the pH sensor in the Monitor to calibrate drop down menu 3 Enter the actual pH value in the Enter reference pH field 4 Click Calibrate 5 Close the Calibration dialog xuri Cell Expansion System W25 User Manual 29 0646 22 AA 189 5 Operation 5 4 Prepare cu
120. g bioreactor etc Enter correct net weight and press tare If correct weight is unknown remove all load Cellbag biore actor etc from tray and tare zero the scale Mechanical obstruc tions to the rocker Check that the tray is firmly locked in position Check that no tubing is pulling on the Cellbag bioreactor during rocking Check that the movement of the rocker is not obstructed Scale needs calibration Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Calibrate the scale See Section 6 1 Calibrations on page 201 If needed contactGE Healthcare ser vice personnel 211 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU 7 3 Xuri Cell Expansion W25 CBCU General Error symptom Status LED is blinking red Possible cause An internal error has occurred but the CBCU is still operating Corrective action Check any warning message and follow the instruc tion If problem persists contact GE Healthcare service personnel Status LED shows a steady red light An internal error has occurred and the CBCU is not operating Check any warning message and follow the instruc tion If problem persists contact GE Healthcare service personnel CAN indicator LED is blinking Gas flow Error symptom An internal error has occurred and the CBCU is not operating proper ly Note This is normal the first seconds during power up Possible cause C
121. g control 253 Media control 286 O mix control 259 pH control Acid Base mode 274 pH control CO2 mode 270 PH control transition de lay 275 oH measurement 267 Pump control 262 Rocking control 250 P Perfusion mode Principles 135 DH Control mode Control scheme 118 Reading cycle 117 Start reading 117 Phases Syntax errors in 230 OH control Acid Base control mode 124 CO2 control mode 122 Transition delays 127 pH control settings Acid base control mode 271 CO2 control mode 269 general 268 transition delay 275 OH measurement principles 115 pH mesurement settings 266 DH sensor Connect 156 Description 34 PID parameters CO2 control 122 O2 control 129 Tuning 122 129 PI parameters Acid Base control 124 Tuning 124 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Index Power switch 17 Prepare for tilt 23 Prepare the system Start UNICORN 170 Process picture 193 Pump Description 29 Status LEDs 30 Troubleshooting 226 Pump 25 specifications 240 Pump control general settings 260 manual run settings 260 Pumps Emergency stop 138 R Reading cycle 117 Reports Add objects to a report 106 Alignment toolbar icons in Customize Report 108 Change the page setup 104 Create a new report for mat 100 Customize Report general toolbar icon functions 102 105 Customize Report toolbar command buttons 101 Edit an existing report for mat 111 Generate and pri
122. he Phase Properties tab 68 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 5 4 Text instructions Step Action 2 Select the variable to be edited if not already selected Do one or several of the following as appropriate Edit Variable Gastlow setpoint Heating setpoint pH reading cycle time Rocking Angle Rocking Speed Hew name C02 setpoint Set visible in details only Rename Delete Close e Type in a new name in the New name field and click Rename e Check the Set visible in details only if the variable should be a detailed variable Uncheck the box to set it to a normal variable e Click Delete to delete the variable Confirm that you want to delete the variable in the dialog that appears 3 Click Close to close the dialog Edit a variable using the VAR button in the Instruction Box The table below describes how to edit a variable using the Instruction Box Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 69 3 The UNICORN software 3 3 Methods in UNICORN 3 3 4 Text instructions Step Action 1 Select the instruction containing the variable to be edited in the Text Instruc tions area Result The parameters for the instruction are shown in the Instruction Box E Main 0 00 Base Time 0 00 Phase Method Settings 0 00 Base SameAsMain 0 00 Rocking 20 0 Rocking Speed RPM 6 0 Rocking Angle o 12 0 o 5
123. he CO setpoint for the next control cycle The CO setpoint is by default limited in the range 0 0 to 15 0 This range can be narrowed using the manual instruction pH control CO2 PID parameters and cycle time The control dynamics differ between the different bag sizes and are dependent of the rate of the gas flow The PID parameters and the cycle time are therefore automatically computed with respect to the bag size and gas flow set point To optimize the control behavior the automatic PID parameters are not the same when approaching a new setpoint as when keeping the setpoint The automatic D parameter is always zero Tuning PID parameters and cycle time If further tuning of PID parameters and or cycle time is needed follow these steps Step Action 1 Open Tools Customize and select the tab Run Data Groups Select the Auto pH CO2 P Auto pH CO2 I and Auto pH CO2 D and click OK Result The automatic PID parameters are shown as Run Data 2 Register the values both when approaching the new setpoint and when keeping the setpoint Tip These automatic values are a good starting point for the tuning 122 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 2 pH control 4 2 1 CO2 control mode Step Action 3 Open the manual instruction pH control advanced pH control advanced CO2 and select Manual PID parameter mode and or Cycle time mode and enter the desired values W
124. he Cellbag to make sure it is attached 4 Repeat the above steps to attach the lower Cellbag rod to completely secure the Cellbag bioreactor on the tray 5 Mount the lid on top of the tray NOTICE Keep the Cellbag bioreactor covered with the lid throughout the cultivation to protect the optical sen sors from excessive light exposure 160 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 4 Prepare the pump 5 1 4 Prepare the pump Tubing holder positions The pumphead has two different holder positions to accommodate tubing with different sizes The inner position is for small tubing and the outer position is for large tubing The image below shows the inner position for small tubing Image below shows the outer position for large tubing The wall thickness of the tubing should be 1 6 mm 1 16 The table below specifies the appropriate tubing inner diameters for the two positions xuri Cell Expansion System W25 User Manual 29 0646 22 AA 161 5 Operation 5 1 Prepare the system 5 1 4 Prepare the pump Tubing inner diameter Tubing inner diameter Tubing holder position mm 0 5 1 50 Inner NOTICE If the tubing holder is in its inner position when larger tubing is used flow and tubing life will decrease If the tubing holder is in its outer position when smaller tubing is used there is a risk for rupture and wand
125. he scale with the weight of the content in the Cellbag entered as Net Weight Temperature control is not functioning or dis played temperature appears to be incorrect 210 Temperature sensor is not in contact with the culture medium Make sure that there is enough culture medi um in the Cellbag bioreactor to cover the temperature sensor also when the rocker is rocking Check that no crease with resulting air pocket has formed on the Cellbag bioreactor film covering the sensor Make sure that the pH or DO cables are not in contact with the temperature sensor Temperature sensor needs calibration Calibrate the temperature sensor See Section 6 1 Calibrations on page 201 If needed contactGE Healthcare service personnel Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Weight Error symptom No weight reading Possible cause Scale does not function properly 7 Troubleshooting 7 2 Xuri Cell Expansion System W25 rocker Corrective action Contact GE Healthcare service personnel Incorrect weight dis played The weight distribution on the rocker feet is not even Use the tare functionality and even the weight distribution according to the instructions in Sec tion 5 4 4 Addition of culture medium on page 185 The rocker is not placed ona horizontal surface Place the rocker on a horizontal surface The tare was not done before adding the load Cellba
126. hen elaborating with a manual parameter remember that increasing the parameter will reduce the integrating effect and vice versa Manual instructions System 2 Instructions p Parameters for pH control advanced C02 a Aone ser PID parameter mode A Heating Gas flow Auto Manual m CO2 mix Manual P 0 00 10 00 02 mix dpi r pH sensor 0 357 Soe Manual 1 100000 pH control advanced m r pH control advanced C02 L 4100 E DO sensor Manual D 0 10000 DO control a DO control advanced rl OV a Save result as L Browse Auto update of parameters during run Click Execute Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 123 4 System control description 4 2 pH control 4 2 2 Acid Base control mode 4 2 2 Acid Base control mode Introduction At every control execution in Acid Base control mode a PI regulator computes the pump run time The pump run time Is given in percent of the cycle time in the range 75 to 75 where the negative part corresponds to the acid pump and the positive part corresponds to the base pump When using the CO2 Base control scheme the control output range is restricted to 0 to 75 to only allow base In the Acid Base control mode the flow rates of the acid and base pumps are adaptive to the actual control case They are automatically and individually set by the software based on the weight of the
127. i Result The Select Method dialog opens In the Select Method dialog browse to the required method and click OK Result The method is added to the method queue Note For reasons of system compatibility the individual methods should be saved for the system on which they are queued xuri Cell Expansion System W25 User Manual 29 0646 22 AA 83 3 The UNICORN software 3 3 Methods in UNICORN 3 3 7 Method queues Step Action 6 Select a Start Condition for the method from the drop down list Note The first Method for the first System will always have its Start Condition set to At queue start Available Start Conditions are At queue start The method will begin at the start of the method queue Only available for the first method for each system Immediately after the previous method has ended The method will start when the previous has ended on the queue for that system Wait The method will start after a specified Wait time has elapsed since the previous method in the queue for the system has ended A separate di alog will open where the Wait time can be specified in Hours and Min utes The delay time will be shown in the Method Queue dialog once entered At ready command The method will start when a Ready instruction in a method on another system has been executed Using this start condition it is possible to connect methods running on different systems A separate dialog will open where the Syste
128. ication Tubing Inner diameter Outer diameter Length Tygon E3603 1 8 3 2 mm 1 4 6 4 mm 157 5 400 cm Silicone 3 16 4 8 mm 3 8 9 5 mm 7 9 20 cm Connectors Inner diameter Reducer connector gas tubing 1 8 to 3 16 3 2 mm to 4 8 mm Connector CBCU 1 8 3 2 mm 246 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 7 Auto calibration 8 7 Auto calibration Description These are the steps in auto calibration Step Action 1 2 10 Ii A message is sent to inform the user that auto calibration has started The pumps stop The system pauses for 15 seconds The weight is measured using averaging during 30 seconds The harvest waste pump runs at the perfusion flow rate setpoint for 2 to 30 minutes depending on the setpoint value The system pauses for 15 seconds The weight is measured using averaging during 30 seconds A calibration command is sent to the harvest waste pump estimating the pumped volume by the measured weight difference between the second and the first weight average The feed pump runs at the perfusion flow rate setpoint for 2 to 30 minutes depending on the setpoint value The weight is measured using averaging during 30 seconds A calibration command is sent to the feed pump estimating the pumped volume by the measured weight difference between the third and the second weight average xuri Cell Expans
129. icture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Acid Base control mode output data Description View from PP RD CD Auto Acid Base P The P parameter used if PID parameter mode is set to Auto Auto Acid Base The parameter used if PID parameter mode is set to Auto Auto pH acid base cycle The cycle time used if Cycle time mode is set to Auto time pH acid base regulator The output of the acid base control expressed as pump output run time in of the cycle time Negative values shall be interpreted as acid positive values as base Total acid The accumulated volume computed by the pump based on the estimated flow and run time Total base The accumulated volume computed by the pump based on the estimated flow and run time 1 This run data is not specifically connected to the acid or base role Instead this run data is connected to a specific pump head Look at the run data from the pump head of interest Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable 274 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 8 pH control Transition delay settings The following table shows the transition delay settings used for pH control Setting Description View from P
130. icture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable CO2 control mode output data Description View from PP RD CD Auto pH CO2 P The P parameter used if PID parameter mode is set to Auto Auto pH CO2 The l parameter used if PID parameter mode is set to Auto Auto pH CO2 D The D parameter used if PID parameter mode is set to Auto Auto pH CO2 Cycle time The cycle time used if Cycle time mode is set to Auto Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable 270 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Acid Base control mode settings Setting The following table shows Acid Base control mode settings used for pH control Description 8 Reference information 8 8 Control settings 8 8 8 pH control Set from PP MI View from PP RD CD Acid The molarity of the acid used expressed Set pH control molarity as equivalent of HCI concentration tings acid mol L If the strength pKa of the se Cellbag lected acid differs from HCI try to esti pumps mate equivalent molarity or see manu Acid al for approximate conversion values pump for other acids The higher molarity of A B C the selected acid the less amount of or D acid will be added by the pH control and vice versa
131. iecsescssisedivenierkeadeathas Guceensvindiaianlipveibuesiiatiionnsiaire 53 555 VOGT A COGS enre cares oriesvieen steams coin E EO emai 57 35 4 TERURE O UCU ONG orra EE E E TEATER en en nT ne tea ere ere 64 5 5 5 Save COO rero s sa R E ET iene ata 72 330 COU T aa E EEEE S del ited ecards ele debs sed AAO 76 55 7 Meinod OO CUS estos cers cori rE N TNE 82 24 Evalugtion in UNICORN essensies E E 86 34 1 EAO O e E E A E sana niece 87 3 4 2 Open orna View TESUNS eriarstina REE LNE EAER 89 34 3 R n documentation sssssssssnisssssnenanaairii o n nS iie on 93 3 4 4 Generate and print a predefined report format eessescssesssessssescssesssecsseessecssessssessssecsseees 98 3 4 5 Create o new report TOMOU snssnasorasisenesnisiassiniiann nina ube rbestiatasestts 100 3 4 6 Edit an existing report TOMOT sssiiirirseirssisssiisrstirrenistintistrednsrsitairaisasniseaki tateitei inainta 111 4 System control description eeseesesssesseseerseeserseeseeseesserseesersceseesceesessesseeseeseeseeeses 113 Al OPRGNG DO measurement and COMMON ssesissiscsssssssssseristndisssiuckoskinnsosiosestoi astiune 114 4 1 1 pH and DO measurement principles sccscscissciessicvcsnssssessestsvevtssstadstesativesievseses esaesensssetcctssecess 115 412 oN aonad BLOB cf 0 4 9 eresio aE T IN AAAA O OOOO OESE 117 4 1 3 OH ond DO contral ONE DOS areara a anea EE ENOO 118 e PEON A A A E E E 121 4 2 1 COZ canol MOGE espiran a E EE trtrertre tie erent 122 Ace Acid Base control Mode siccss
132. ight decreases Pump calibration 134 Since the feed pump flow is regulated to keep the momentary weight setpoint no precise calibration of the feed pump is needed Following the momentary weight setpoint will automatically give the correct feed rate anyway Setting the right tube inner diameter will be enough Calibrating the pump before running media addition has the advantage of getting the accumulated feed volume more precise Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 4 Media control 4 4 2 Perfusion 4 4 2 Perfusion Introduction In perfusion mode the feed pump will add medium at a desired flow and the har vest waste pump flow is controlled by a regulator to keep the given weight setpoint The true feed flow is dependent of how well the feed pump is calibrated To make the calibration more convenient and to compensate for tubing wear out auto calibration of the feed and harvest waste pumps is enabled by default Perfusion principles When starting the media control the feed or harvest waste pump will first run alone until the weight setpoint is reached When the setpoint is reached normal perfusion starts and both pumps will run If the weight setpoint is changed while in normal perfu sion one of the pumps will run alone until the new setpoint is reached The figure below shows a run where the control is started with the measured weight below the weight setpoint Af
133. ination of acid and base as actuators Se lect CO2 for using only CO as actuator PH setpoint The pH target value Settings pH OH control y Y Y general Deviation Used to select if the deviation pH control Y alarm alarm should be active or not general Allowed devia A parameter that defines how Settings pH pH control Y tion up much above the pH setpoint the general pH reading is allowed to be without triggering the deviation alarm Allowed devia A parameter that defines how Settings pH pH control Y tion down much below the pH setpoint the general pH reading is allowed to be without triggering the deviation alarm 268 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 8 pH control Setting Description Set from View from daa gt Oe D Deviation A parameter that defines for pH control alarm delay how long the pH reading can be general outside the allowed deviation limits before the deviation alarm is triggered Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable CO2 control mode settings The following table shows CO2 control mode settings used for pH control Setting Description Set from View from MI PP RD Max CO2 The maximum CO concentration pH control CO2 y concentra set
134. ing 4 Transfer the desired volume of inoculum into the bag using a pump or gravity flow 192 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 5 Perform cultivation 5 5 2 Monitor and control the cultivation 5 5 2 Monitor and control the cultivation System status You may follow and control the ongoing run in the System Control module The current system status is shown in the System state panel in the Run Data pane For example it may state Ready Manual Run or Method Run om Tana x Tube s i O B bia Fle 1 1G haia E OE kajaa l aa er hosh Ccann s Connected im conte Dentredled by Pairi pOIEPI LT Process picture The Process Picture displays the process parameters during a run During a run real time data from monitors are displayed The Process Picture is shown in the illustration below The buttons available depend on the system configuration 00 54 0 a S Pi i A 02 02 10 65 Rocking 30 on gt Temp Media control pH 6 83 S co2 0 0 B Gas flow 1 00 limin Samp ng Smin jA oe conie pi 6 83 A a n bag pumps anase Cellbag Weight Rocking 4 me Cellbag 2L weight 0 72 9 3 N Loa feed 2 0 49 gt By Pump 25 24 Harvest 30 341 gt Ea ees Purp 75 25 The button including its frame will have a different color depending on the state of the function Color indication is applied as shown in the t
135. ining syntax errors is marked in the method outline with an error symbol a white cross on a red circular background The table below de scribes some solutions to syntax error problems Problem description The method instructions do not correspond to the components you have chosen for your system Solution Check your system components under System Proper ties in the Administration module and that the correct instrument configuration is selected Syntax errors are not corrected by changing the component configuration Close and reopen the method Syntax errors appear because the method was connected to the wrong system That is the instrument configuration of the sys tem is incompatible with the method Edit the method so it can be run on the currently chosen system Save the method for a system that has all compo nents installed Note The red instructions must be replaced or removed Reselect the required component under System Properties in the Administration module if the component is actually present on the system Re open the method and replace the red instructions with the corresponding instruction for the added component Syntax errors appear because the system s instrument configuration has been updated with a new instrument configuration that differs in the instruction set 230 Select the red instruction and either delete it or replace it with a corresponding instruction if av
136. instructions Step Action 4 Click OK 5 To save the changes click Change in the Instruction Box Result The text instruction is updated Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 71 3 The UNICORN software 3 3 Methods in UNICORN 3 3 5 Save a method 35 5 5 Introduction Save a method Methods and phases are saved in the UNICORN database Individual edited phases may be saved to the Phase Library for later use in other methods on systems having the same instrument configuration and component config uration Save a method 72 The table below describes how to save a method in UNICORN Step Action 1 e Click the Save the Method icon ka or e select File Save or File Save As Result e lf the method has been named and saved previously the changes are saved immediately If not e the Save As dialog opens Proceed with steps 2 4 below Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 5 Save a method Step Action 2 Browse for an appropriate folder or create a new one Last modified Created by Bridget BT 2012 1 225 Default Charlie 25 1 efault H Charl 57187 2012 1 23 1 Defaul shine 3 U efault gt Chri i 6 18 2012 1 23 0 Detaul z AuBielo BY 1a 2012 1 245 Default H David E 57187 2012 1 23 35 Default David T 5187 2012 1 232 Default E DetaulktH ome g 25r2012 1 07 1
137. instructions below to verify that the temperature control is functioning cor rectly Step Action 1 Switch on the client computer Z Switch on the rocker and wait for the system to initialize x Start UNICORN and connect to system 4 Attach a Cellbag bioreactor and fill with at least the minimum volume of liquid Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 145 4 System control description 4 6 System verification Step Action 5 10 11 12 Make sure that the Temp button in the Process Picture displays the ambient temperature Make sure that the rocking is on Heat will not be applied to a motionless rocker to avoid local overheating in the Cellbag bioreactor Start the heating by clicking the right hand side of the Temp button Result The heating is activated and the Temp button gets green Temp 21 7 Enter the temperature 40 0 in Settings Temp Setpoint in the Process Picture Click OK Result The temperature starts to increase to the set temperature value CAUTION To avoid overheating do not operate the heater with out liquid in the Cellbag bioreactor on the tray Select the Chart tab and make sure that the temperature curve shows the increase in temperature If the temperature control appears not to function correctly refer to Sec tion 7 2 Xuri Cell Expansion System W25 rocker on page 208 Stop the heating by clicking the right hand side of the Temp but
138. ion Possible cause This is normal and may be caused by differ ences in temperature atmospheric pressure and composition of the gas 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Corrective action Check that the correct calibration values have been entered If problem persists perform an offset calibra tion Note If for example 5 CO is added to air flowing through the Cellbag the DO will be reduced with approximately the same percentage value Incorrect DO reading Media control Error symptom A gas bubble may be trapped on the DOOPT Il sensor Possible cause Tap the DOOPT II sensor from the underside to re move the bubble This may require forceful manip ulation and the Cellbag bioreactor may need to be partly disconnected from the tray Corrective action Media control is not starting Some of the criteria for starting the Media con trol are not fulfilled Read the message in the pop up dialog If possible take action according to the information and try again Media control is inac tive Rocker is in sampling mode None The media control is inactive while the rocker is in sampling mode Some of the criteria for running the Media con trol are no longer ful filled Read the message in the pop up dialog If possible take action according to the information and make sure that the control turns active again In Perfusion mode only the
139. ion System W25 User Manual 29 0646 22 AA 247 8 Reference information 8 8 Control settings 8 8 Control settings In this section 248 This section lists the different control settings that can be made in UNICORN This section contains the following subsections Section See page 8 8 1 Rocking control 249 8 8 2 Heating control 252 8 8 3 Gas flow control 254 8 8 4 CO mix control 256 8 8 5 O mix control 258 8 8 6 Pump control 260 8 8 7 pH measurement 265 8 8 8 DH control 268 8 8 9 DO measurement 2 76 8 8 10 DO control 279 8 8 11 Media control 284 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 1 Rocking control 8 8 1 Rocking control Rocking control settings The following table shows general settings used for rocking control Setting Description Set from View from PP MI PP RD Speed The number of rocking cycles Settings Rocking Yy Y setpoint per minute Rocking Angle The maximum angle between Settings Rocking Y Y setpoint the tray and the horizontal plane Rocking during a rocking cycle Stop angle The angle between tray and Settings Rocking Y Y horizontal plane the tray will Rocking settle at when the rocking is stopped At a positive stop angle the tray will lean towards the front side of the rocker and vice versa Sampling The number of minutes the Settings Rocking Y mode rocker will stay in sampling Rocking
140. ioreactor 9 Disconnect the tubing and reconnect fresh culture medium to refill the Cellbag bioreactor Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 5 Perform cultivation 5 5 2 Monitor and control the cultivation Step Action 10 Open the inlet and outlet gas filter clamps 1l Start the rocking by clicking the right hand side of the Rocking button in the Process Picture 12 Start the gas flow by clicking the right hand side of the Gas flow button in the Process Picture Perfusion culture During perfusion culture cell free harvest waste is withdrawn and fresh medium is added continuously Requirements e Cellbag bioreactors with internal perfusion filter or connected to external retention filters e Harvest waste and feed pump See Section 4 4 2 Perfusion on page 135 for information on perfusion mode Contact a GE Healthcare application specialist for advice on setting up perfusion culture xuri Cell Expansion System W25 User Manual 29 0646 22 AA 197 5 Operation 5 5 Perform cultivation 5 5 3 End cultivation 5 5 3 End cultivation End cultivation and harvest To end the cultivation and harvest follow the instructions below Step Action 1 Prepare the harvest vessel 2 Click the stop button in the toolbar in System Control to stop the run 2 System Control File Edit view Manual System Tools Help llelo e eA When asked if you want to end the run click OK
141. is parameter has no effect Start acid pump Acid pump speed The rom at which the acid pump will run Settings Cell bag pumps 1 Start acid pump Base pump mode If Limited is selected the pump will run as long as given by Base pump duration If Continuous is selected the pump will run until the base pump is stopped Always Contin uous when started from PP Start base pump Base pump duration The time the base pump will run if Base pump mode is set to Limited Else this parameter has no effect Start base pump Base pump speed The rpm at which the base pump will run Settings Cell bag pumps 1 Start base pump Feed pump mode If Limited is selected the pump will run as long as given by Feed pump duration If Continuous is selected the pump will run until the feed pump is stopped Always Contin uous when started from PP Start feed pump Feed pump duration Feed pump speed The time the feed pump will run if Feed pump mode is set to Limited Else this parameter has no effect The rom atwhich the feed pump will run Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Settings Cell bag pumps 1 Start feed pump Start feed pump 261 8 Reference information 8 8 Control settings 8 8 6 Pump control Setting Description Set from View from PP RD Harvestpump If Limited is selected the pump Always Contin
142. issessenissseaiesasernsnercrcesesesersspsiaonniprioaviesvigsieaarsineetternenasatanmnrireninne 124 42 3 DACON TOTON 6 24 9 11 aE E Pte tee errr ent verte 127 CEG CO ce caterers neta E E A EAN AE E eee eae 128 4 5 1 Oron ONO E eero a E RCT I OEN E EE 129 4 3 2 iS 8 216 T 1416 6 em nn me ets OO E On OTs ONSET PE ie CER Te Te ee T51 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 Table of Contents 4 3 3 DO COTO TROIS TOR delays sessirnar SRN 132 Al Mead CONTO sre A E O E E EEN 133 4 4 1 Lire lao 6 0 O Seem nee een eer tr ea A REE NO OEE eT ee tet ere verre nnn te tet 134 4 4 2 Pe eae nnn ns rey nnn Entei ee vy ee 135 4 4 3 Deviation Glar seess e r AASS 139 4 4 4 Media contona AIOI isea A A 140 4 5 Recommended operating conditions sssssssssssssssssssssssssssssssssssssserssresrsssresrirrrrrrrrrrrerrrerrsrrsess 141 A6 Sytten vernicosa S E 144 E E y E EAE T AA PN AA NA VEA A AE T AT 150 S PE E a a E E E AEA OEE 151 5 1 1 Select tray and Cellbag bioreactor eeecseecssesssessssessssessssecsssessuecsssssssessssecsssecssscsssssssecessecssseessee 152 5 1 2 Prepare pH and DO Senso do gaara ete eeennn ee une eT R te st neues OPE EY ane en TeNT Pete rO Cer 156 ILs Attach the Cellbag bioreactor eeseessesseseseirisesesssssssssssssssrrrresssssssssssssssrssrrrrssssssssssssssssnn 159 5 1 4 Prepare the I casts cacesens costco ectoas face en geechcaccieapatn cemeap E E AAEE 161 5 1 5 OTC CE GS TO VSTE areca
143. ition delays CO2 Base control scheme The CO2 Base control scheme is combining the CO2 and Base control modes It always starts in CO2 mode and can then shift back and forth between CO2 and Base mode as long as the control is on To avoid unnecessary toggling between CO2 and Base mode the transitions between the modes are delayed This delay is called transition delay time The condition for transition must be fulfilled for a certain time before the transition takes place The transition delay times are set automatically and individually by the pH control The values can be overridden with manual values using the manual instruction pH control transition delays CO2 mode Condition Condition pH reading gt pH set point pH reading lt pH set point AND AND CO2 set point at minimum fullfilled for the Base CO2 to base transition delay time mode If the pH CO controller is at its minimum CO setpoint and the pH reading is below the pH setpoint a time counter is started When the time counter reaches the set CO2 to base transition delay time the transition to base mode takes place If the pH base controller is at its minimum output 0 and the pH reading is above the pH setpoint a time counter is started When the time counter reaches the Base to CO2 transition delay time the transition to CO mode takes place Base pump time at zero fullfilled for the Base to CO2 transition delay time Xuri Cell Expansion System W25
144. ition delays See Manual instructions on page 47 The control is using a manually entered tran sition delay time not optimal for the actual case Use the manual instruction DO control ad vanced DO control transition delay Set 02 Speed transition delay mode to Auto See Manual instructions on page 47 DO control is inactive DO reading Error symptom Some of the criteria for running the DO control are no longer fulfilled Possible cause Read the message in the pop up dialog If possible take action according to the information and make sure that the control turns active again Corrective action No DO reading Off is displayed DO reading is not start ed Open Settings DO from the Process Picture and set Reading to On Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 221 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Error symptom DO reading is inactive 0 0 is displayed and the frame around the Process Picture button is orange Possible cause Repeated errors from the the sensor has turned the reading into an inactive state New reading attempts are made with asked read ing frequency until a valid reading is re turned or the reading is stopped Temperature of the cul ture medium has not reached process tem perature Corrective action Read the warning messages for more information Check the possible causes and corrective action
145. just the tubing holder to the correct position for your size of tubing See instructions above 164 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 4 Prepare the pump Step Action 4 Place the tubing between the rotor rollers and the track pressed against the pumphead inner wall 5 Lower the flip top until it clicks into its fully closed position Note The pumping direction is clockwise xuri Cell Expansion System W25 User Manual 29 0646 22 AA 165 5 Operation 5 1 Prepare the system 5 1 5 Connect gas to system 5 1 5 Connect gas to system Gas mix The Cellbag bioreactor requires gas flow to stay inflated and to provide ventilation The CBCU enables different gas mixing possibilities Compressed air or N can be mixed with CO and or O to obtain the desired gas mix Compressed air or N is connected to AIR N2 on the CBCU CO and O are connected to CO2 IN and O2 IN respectively on the CBCU rear panel 166 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 5 Connect gas to system Set up the gas system Follow the instructions below to connect gas to the bioreactor system Step Action 1 Attach the filter heater to the outlet gas filter of the Cellbag bioreactor The image below shows the filter heater mounted on the stand on the Cellbag bioreactor xuri Cell Expansion System W25 User Manual 29
146. l Deviation alarm Used to select if the deviation alarm should be active or not DO control general Allowed devia tion up A parameter that defines how much above the DO setpoint the DO reading is allowed to be without triggering the deviation alarm Settings DO DO control general Allowed devia tion down A parameter that defines how much below the DO setpoint the DO reading is allowed to be without triggering the deviation alarm Settings DO DO control general Deviation alarm delay A parameter that defines for how long the DO reading can be outside the allowed deviation limits before the deviation alarm is triggered Xuri Cell Expansion System W25 User Manual 29 0646 22 AA DO control general 219 8 Reference information 8 8 Control settings 8 8 10 DO control Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable O2 control mode settings The following table shows O2 control mode settings used for DO control Setting Air source Description If set to Compressed air the output range of the DO O2 mode controller is set to 21 50 If set to N2 the output range of the DO O2 mode controller is set to 0 50 Select air source View from PP RD CD PID param eter mode When Auto is selected the syste
147. lder name system xa UNICORN Database Eric Methods Method Systema El Method Systema Results 3 To open the method e Click the Open button located in the toolbar of the Method Navigator pane Open or e double click the selected method or e Right click on the method name and select Open from the context menu Result The method is opened and displayed in the Method Outline pane with included phases You can continue to edit the phases of the method using Phase Properties or manually text edit the method in the Text Instruc tions pane 58 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 3 Work with methods Add a user defined phase to the method outline using drag and drop The table below describes how to add a user defined phase to the method outline using drag and drop Step Action 1 Select the User Defined phase in the Phase Library pane and drag and drop the phase to the requested position in the Method Outline pane Result The phase is included in the method at the requested position 2 When the User Defined phase has been added to the Method Outline the phase name is enabled for editing User Defined iT F User Defined iT Type a name for the phase and press the Return keyboard key Note The User Defined phase is marked with the letter T meaning that it is text edited This phase contains only Base and End_Block inst
148. llbag bioreactor Overinflation Error symptom Possible cause Corrective action Cellbag bioreactor ap Too high gas flow Check that the gas flow to the Cellbag bioreactor pears to be overinflat does not exceed 1 Ipm or that the Fast fill function ed is turned off Faulty relief valve Check that gas flows through the pressure control valve e Attach a short length of tubing to the outlet valve Place the tubing end into water to 1 cm depth Result Bubbles indicate flow If no bubbles are observed remove the pres sure control valve The pressure control valve may be blocked and removing It may allow continued operation Outlet filter is closed off Make sure that the outlet filter is not closed off or blocked or blocked If foam is clogging the outlet filter take action to reduce the foam Decrease the rocking speed and or rocking angle add antifoam and or slightly increase the gas flow rate Faulty Cellbag bioreac If the Cellbag bioreactor continues to overinflate tor transfer the contents to another Cellbag bioreac tor Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 227 7 Troubleshooting 7 5 Cellbag bioreactor Underinflation Error symptom The Cellbag bioreactor appears to be underin flated Possible cause Too low gas flow Corrective action Check that there is sufficient gas flow to the Cellbag bioreactor Inlet gas supply incor rectly connected
149. lly interact with an ongoing run using Manual instructions Follow the instructions below to perform manual instructions Note It is also possible to interact with the system manually directly from the Process Picture Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 47 3 The UNICORN software 3 2 System control Step Action 1 In the System Control module e select Manual Execute Manual Instructions or e use the shortcut Ctrl M Result The Manual instructions dialog opens Instructions Heating Heating Enable heating Disable heating Gas flow CO mix 02 mix pH sensor pH control pH control advanced DO sensor sO 2 In the Manual instructions dialog e Click the symbol to show the instructions for the instruction group that you want to modify e Select the instruction that you want to modify e Enter the new values for the instruction 3 To execute several instructions at the same breakpoint select and edit an instruction and click Insert Repeat for several instructions Note To update parameter fields during run check the Auto update box 4 To perform the instructions click Execute Run data The Run Data pane shows the current values of for example rocking motion pH mea surement and accumulated time Since there are many Run Data the view can be cus tomized To see more information select View Run Data and e right click in the Run Data pane
150. ltivation 5 4 5 Equilibrate the culture medium Step Action 10 1 Select Settings DO in the Process Picture 2 Enter the desired values for e Control e Setpoint e Alert deviation alarm interval 3 Click OK 4 Repeat step 1 to 3 for the pH parameters aa Turn on DO and pH control by clicking the right hand side of the relevant buttons in the Process Picture 12 Make sure that the key culture parameters pH DO and temperature are stable before inoculation 190 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 5 Perform cultivation 5 5 Perform cultivation In this section This section describes the basics of performing a cultivation During the cultivation key parameters are monitored and the settings can be adjusted This section contains the following subsections Section See page 5 5 1 Inoculation 192 5 5 2 Monitor and control the cultivation 193 5 5 3 End cultivation 198 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 191 5 Operation 5 5 Perform cultivation 5 5 1 Inoculation 5 5 1 Inoculation Instructions Follow the instructions below to inoculate the Cellbag bioreactor Step Action 1 Make sure that the inlet tubing and the tubing connected to the inoculum container is clamped 2 Connect tubing from the inoculum container to the inlet tubing using e g tube fusing equipment or a ReadyMate connector 3 Unclamp the inlet tubing and inoculum container tub
151. m and Method can be chosen Method Queue Number of included systems 1 v System v Method Start Condition 1 Methodi lza At queue start 2 Method2 ee Immediately after the previous method has ended Insert Row 3 Method3 Wait Oh 30min after the previous method has ended T Repeat steps 4 to 6 to add further methods to the Method list for each re quired system 84 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 7 Method queues Step Action 8 Click Save or Save As to save the completed method queue Note An error dialog will be displayed if any of the methods are incompatible with the system on which they are queued Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 85 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 Evaluation in UNICORN In this section This section describes how to use the Evaluation module to evaluate the results of a run For information about dialogs in the Evaluation module that are not described in this manual refer to the online help This section contains the following subsections Section See page 3 4 1 Evaluation 87 3 4 2 Open and view results 89 3 4 3 Run documentation 93 3 4 4 Generate and print a predefined report format 98 3 4 5 Create a new report format 100 3 4 6 Edit an existing report format 111 86 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN softwa
152. m will automatically set the PID parameters When Manual is selected the system will use the manual PID parameters set by this instruction DO control advanced 02 A hand symbol is shown if set to Manual Manual P The P parameter used if PID parame ter mode is set to Manual DO control advanced 02 Manual The l parameter used if PID parame ter mode is set to Manual Note that increasing the I parameter will reduce the integrating effect and vice versa DO control advanced 02 Manual D The D parameter used if PID parame ter mode is set to Manual DO control advanced 02 Cycle time mode When Auto is selected the system will automatically set the cycle time When Manual is selected the system will use Manual cycle time DO control advanced 02 A hand symbol is shown If set to Manual Manual cy cle time 280 The cycle time used if Cycle time mode is set to Manual DO control advanced 02 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 10 DO control Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable O2 control mode output data Description View from PP RD CD Auto DO 02 P The P parameter used if PID parameter mode is set to Auto Auto DO 021 Th
153. me height Adjusts the selected objects to the same height as the highlighted reference object Note The Make same size and Make same width functions can only be used to resize the width of charts free text and picture objects Save the report format The table below describes how to save the finished report format Step Action 1 e Choose File Save or e Click the Save icon g Result The Save Report Format dialog box opens d e Type a name for the format e Select if you want to save the format for global use e Select if you want to save the format as default Note The name for the default format will automatically be changed to DEFAULT xuri Cell Expansion System W25 User Manual 29 0646 22 AA 109 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Step Action 5 e Click OK to save the format 110 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 6 Edit an existing report format 3 4 6 Edit an existing report format Introduction This section describes how to edit an existing report format Note By clicking the Preview Edit mode button you can toggle between a print preview of the report and an editing mode The editing actions are only avail able in the Edit mode Edit a saved report format The table below describes how to edit a saved report format in the Evaluation module Step
154. mpling connector 194 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 5 Perform cultivation 5 5 2 Monitor and control the cultivation Step Action 4 Wipe the top of the sampling con nector with 70 alcohol or equiv ll alent 5 Attach a sterile disposable syringe with luer connector onto the sam pling connector j 6 Release the tubing clamp and withdraw a sample into the syringe 7 Remove the syringe and wipe the top of the sampling connector again with 70 ethanol and replace the cap 8 Pinch the sampling connector tubing a few times to ensure that any liquid in the tubing drains back into the Cellbag bioreactor 9 Close the tubing clamp Sampling mode duration is set from Settings Rock ing Sampling Pause in the Process Picture Scale up cultivation Cellbag bioreactors have a large range in operating volume This allows efficient scale up and eliminates tedious sequential transfers Start at low volume and add fresh medium to the Cellbag bioreactor as the cells grow Up to a 1 10 expansion is possible in one single bag Example of an inoculum expansion sequence is described below xuri Cell Expansion System W25 User Manual 29 0646 22 AA 195 5 Operation 5 5 Perform cultivation 5 5 2 Monitor and control the cultivation Step Action 1 Start with 300 mL medium in a 2 L Cellbag bioreactor Add inoculum 2 When cells reach approximately 3 x 10 cells mL add culture medium up to
155. n several charts from different re sults using File Open to Compare Refer to Online help for furhter information Note When working in a network environment it is not possible to edit the same result file from two different locations simultaneously If the result is already open at another network workstation you can only open it in read only mode A message similar to the illustration below will open xuri Cell Expansion System W25 User Manual 29 0646 22 AA 89 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 2 Open and view results Evaluation The result is locked for editing by Markus at computer SEHC2GELY2I0 Do you want to open a Read Only copy Highlight or select a curve You can highlight or select an individual curve in the chart The table below describes the differences hold your mouse pointer a pop up box will display the curve name over a curve segment hold your mouse pointer the curve and the short line segment in front of the curve over a curve name name become bold click a curve segmentor the Y axis shows the values for this specific curve a curve name Insert a vertical marker The vertical marker is used to measure the values for a specific curve position Right click in the chart and choose Vertical marker Move the marker along the X axis and read the X axis and Y axis values of the selected curve in the box in the top left corner of the chart Weight Chert tph J
156. nformation 8 4 Xuri Cell Expansion W25 CBCU semi wetted materials 8 4 XuriCell Expansion W25 CBCU semi wetted materials This list covers semi wetted materials i e materials that get into contact with the gas flow from gas source to gas outlet in Xuri Cell Expansion W25 CBCU Those parts of the flow path that do not go to the outlet such as connections to pressure sensors and flow path to O and CO sensors are not included For some of the materials listed the ASTM D1418 standard practice for rubber and rubber lattices nomenclature abbreviation is written in parenthesis Gas distribution block pressure relief valve Material Aluminum Flow regulators pressure relief valve Brass Flow restrictors pressure relief valve outlet connector Stainless steel O rings gaskets Pressure relief valve Ethylene Propylene Terpolymer EPDM Nitrile NBR Check valve Fluoroelastomer FKM Tubing Silicone VMQ Tubing connectors Flow regulators Polyamide Fluoroelastomer FKM FPM Check valve Polypropylene Tubing connector between Xuri Cell Expansion W25 CBCU and gas source Animal free polypropy lene resin Tubing connector between Xuri Cell Expansion W25 CBCU and Cellbag bioreactor Animal free polypropy lene resin Gas distribution tubing between Xuri Cell Expansion W25 CBCU and gas source Tygon E3603 Gas distribution tubing between Xuri Cell Ex
157. nged the changes will be saved with the method 2 e click the Save Phase button below the Method Outline pane r O e select Phases Save Phase Insert Phase From Library Save Phase k or e right click the phase and select Save Phase Renae Delete Move Up Move Down ut Copy 74 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 5 Save a method Step Action Result The Save Phase to Phase Library dialog opens Save Phase to Phase Library Phase name For system System 2 Global Personal 3 e Type a Phase name or e Choose a phase from the Phase name drop down list This phase will be replaced by the phase with the new settings 4 In the For system field the system that was selected when the current method was set up will be displayed by default To save the phase for another system choose the appropriate system from the For system drop down list Note Only systems using the same instrument configuration and component con figuration as the system that was selected when the current method was set up will be displayed in the For system field 5 e Select if the phase shall be Global available for all users or Personal for your own use only Result The phase is saved and is available in the Global Phases or Personal Phases panel of the Phase Library pH 3 io 12
158. ns pumphead 2A to Acid System settings Set role Base Feed or Harvest Waste Pump 25 2A role Pump 25 2B Assigns pumphead 2B to Acid System settings Set role Base Feed or Harvest Waste Pump 25 2B role Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Note When sending one system setting all system settings are sent Therefore check the role assigned to all pumpheads before sending a role command to ensure not to have conflicting roles assigned to the different pump heads Settings for manually running the pumps The pump settings in this section are only valid when the pumps are started manually When the pumps are used by pH or media control these settings have no effect Setting the tube diameter and acid base molarity is described under pH and Media control settings 260 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Setting Acid pump mode Description If Limited is selected the pump will run as long as given by Acid pump duration If Continuous is selected the pump will run until the acid pump is stopped 8 Reference information 8 8 Control settings 8 8 6 Pump control Set from Always Contin uous when started from PP Start acid pump PP View from D Acid pump duration The time the acid pump will run if Acid pump mode is set to Limited Else th
159. nt a prede fined format 98 Save a new report for mat 109 Save a report in PDF for mat 99 Result files Open a result 89 Rocker Description 16 Filter heater 25 Front view 17 Function verification 144 Rear view 19 Rocking parameters 16 Troubleshooting 208 Rocker specifications 239 Rocking angle Index 290 Operating conditions 141 Rocking control settings 249 Rocking motion Operating conditions 142 Rocking speed Operating conditions 141 Run start 177 Run documentation Documentation tabs de scription 94 Print 93 Save the used method as a new method 97 Search for log entry text 96 View 93 S Safety notices 9 Scouting Change variable values 79 Overview 76 Scouting scheme 76 Snapshots Take a Snapshot of curve values in a result 92 Software overview 36 software modules 37 Speed step 131 Start protocol Set up start protocol 61 Status LED CBCU 27 Status LEDs Pump 30 Syntax errors In methods 230 In phases 230 In text instructions 230 System Components 230 Instrument configura tion 230 System Control module curves 46 description 42 icons 45 manual instructions 47 process picture 193 run data 48 system settings 47 System properties edit a definition 174 System recommendations computer specifica tions 242 System settings 47 System specifications 237 T Temperature control Function verification 145 Text editing Help texts 64 T
160. nt labeled Xuri Cell Expansion W25 CBCU Control unit for gas mixing pH and DO control Pump Xuri Cell Expansion System W25 Pump Refers to instrument labeled Xuri Cell Expansion W25 Pump Rocker Xuri Cell Expansion System W25 Refers to instrument labeled Xuri Cell Expansion System W25 Bioreactor system The entire system including rocker CBCU and pump together with Cellbag bioreactor and filter heater UNICORN The software used for controlling and monitoring the system 10 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 1 Introduction 1 3 User documentation 1 3 User documentation The table below describes the user documentation for Xuri Cell Expansion System W25 which is available from the Help menu in UNICORN or on the user documentation CD Document Xuri Cell Expansion System W25 Operating Instructions Main contents Instructions needed to install operate and maintain Xuri Cell Expansion System W25 in a safe way In cludes basic UNICORN system control functions Xuri Cell Expansion System W25 User Manual Detailed system descriptions and instructions on how to run maintain and troubleshoot Xuri Cell Expansion System W25 Includes UNICORN system control functions method creation and handling together with evaluation and presentation of data Xuri Cell Expansion System W25 Cue Card Brief instructions providing an overview of how to run the system UNICORN Admini
161. nual settings Here the user has to adapt the parameters manually to the used bag size and gas flow The automatic parameters can be overridden with manually entered values If any of the automatically computed values are overridden a hand symbol see below appears in the Process Picture If hovering with the cursor above the symbol a tool tip informs which parameters are overridden Deviation alarm Inactivation 120 If the reading has been outside the specified deviation limits of the setpoint for a certain time a deviation alarm is activated by default The deviation alarm can be deactivated with the manual instruction pH control general or DO control general After a change of the setpoint the deviation alarm check is inactive until the reading is within the deviation limits of the new setpoint If the interval within the deviation limits is not reached within a time limit computed by the pH or DO control the deviation alarm is triggered In the pH control CO2 mode the time limit is equal to a multiple of 40 of the automatically computed control cycle time In Acid Base mode the time limit is equal to 90 minutes In the DO control 02 mode the time limit is equal to a multiple of 40 of the automatically computed control cycle time In Speed mode the time limit is equal to 180 minutes If any of the conditions for running the pH or DO control is suddenly not fulfilled when running the control the control will t
162. od Editor panes As illustrated below three panes are shown in the Method Editor by default The available User Defined phase is found in the Phase Library 1 and an overview of the phases in cluded in the active method is displayed in a Method Outline 2 Information about the method is presented in the right pane 3 containing the two tabs Phase Properties and Text Instructions Method Editor toolbar icons The table below shows the Method Editor toolbar icons that are referred to in this User Manual Function New Method Opens the New Method dialog where methods can be cre ated Open Method Navigator Opens the Method Navigator where available methods are listed Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 51 3 The UNICORN software 3 3 Methods in UNICORN 3 3 1 Method editor Function Save Saves the active method Print Opens the Print dialog from where a method can be printed Start protocol Opens the Start Protocol dialog where settings for the start protocol can be made Method notes Opens the Method Notes dialog where notes can be added to the method Scouting Opens the Scouting Variables dialog which is used to repeat a series of method runs 52 New method queue Opens the Method Queue dialog Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 3 2 Method creation Predefined method 3 The UNICORN software 3 3 Methods in UNICORN 3 3
163. ontact GE Healthcare service personnel Corrective action No gas flow or fluctuat ing gas flow and a high pressure alarm is acti vated Gas flow has shut down or is restricted due to high pressure caused by blockage in gas tub ing or gas filters e Make sure that all inlet and outlet lines are open e Disconnect the N Air and Gas mix out tubing and locate the blockage Clogged Cellbag biore actor outlet gas filter e Check that the Filter heater functions properly e lf the outlet gas flow is blocked by foam re duce the rocking angle or decrease the rocking speed e f problem persists transfer the contents to another Cellbag bioreactor 212 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Error symptom External gas source is connected but the gas flow shows zero CO mix Error symptom No CO reading or the communication with the CO sensor does not work Possible cause Blocked gas tubing 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Corrective action Disconnect the N2 Air and Gas mix out tubing and locate the blockage External gas source is connected but the tube regulator is not opened Possible cause CO sensor is broken Open the external gas tube regulator Corrective action Contact GE Healthcare service personnel Noimmediate response when CO mix is start ed or when setpoint is changed Display shows
164. or by decreasing the rocking speed and or angle e Excessive foaming occurs If the Cellbag bioreactor is not rigidly inflated Check that the gas flow is sufficient and that the pressure relief valve is functioning Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 143 4 System control description 4 6 System verification 46 System verification Introduction This section describes verification procedures for the rocker function temperature control and gas flow Before verification install the system according to Chapter 4 Installation in the Operating Instructions and Section 5 1 Prepare the system on page 151 System verification is recommended before the first cultivation and if the system has not been used for a long period For more information about system control settings see Section 8 8 Control settings on page 248 Verification of rocker function Follow the instructions below to verify that the rocker operates correctly Step Action 1 Switch on the client computer 2 Switch on the rocker and wait for the system to initialize 3 Start UNICORN and connect to system 4 Start the rocking by clicking the right hand side of the Rocking button Result The rocking is activated and the Rocking button gets green P Rocking 20 rpm 5 Follow these instructions to make sure that the rocking speed functions properly 1 Enter the desired speed in Settings Rocking Speed in the Process Picture Click O
165. or tuning of acid base flow rates To make this automation work the following input data needs to be correct Acid molarity The molarity of the acid and base used shall be entered as equivalents of HCI NaOH concentrations If the strength pKa pKb of the used acid base differs from HCI NaOH try to estimate the equivalent molarity Generally if a high molarity is entered the computed flow will be low and vice versa Knowing that the acid base molarity pa rameters can be used as inverted gain parameters to easily tune the controller if needed The weight measurement shall correspond to the weight of the content of the Cellbag Make sure the scale is tared correctly The inner diameter of the acid and base tubing This data is entered to give the pumps the right relation between commanded flow and rpm A pump calibration will make the relation more precise but entering the tube diameter should be enough for the pH acid base control Acid tube diameter a Compute sotkien Acid flow Weight reading H reading P Pump run time controller pH setpoint Base molarity oac Base flow qd Base base flow pump Weight reading Base tube diameter If the automatically computed flow is lesser or greater than what can be achieved with the entered tube diameter the pH control will inform the user that a smaller or larger tube is needed for optimal control If desired manual flow rates for acid and b
166. ormat that has been defined and saved Should you need to store your reports in an electronic format you can also save them as PDF files This section describes how to do this Generate and print the report The table below describes how to select a format and print the report Step Action 1 e Choose File Report or e Click the Report icon a Result The Generate Report dialog opens 2 e Selecta Format for the report Generate Report Format ol E BIM Test Report Delete Edit Export Import 9 Cen er Note Global report formats are noted by the text Global before the report format name in this list 98 xuri Cell Expansion System W25 User Manual 29 0646 22 AA Step 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 4 Generate and print a predefined report format Action e Click the Preview button to view the report in the Customize Report window and click the Print icon or e Click the Print button Result The Print dialog opens e Selecta printer from the Printer droplist e Select a Print Range all pages or just selected pages e Select a number of copies e Click the OK button Result The report is printed on the selected printer Note Select Edit mode in Customize Report to change the layout You can either print the edited format from this mode and exit Customize Report without saving the changes or save the edits when you exit
167. ort title box and click the Font button to alter the default font if necessary The title is centered immediately above the page contents 5 If you want to have a line under or over the header select the appropriate option in the Layout field 6 e Repeatsteps 3 to 5 on the Footer tab and the subsequent pages Header tab Note All Header and Footer tabs contain the same options You can have all infor mation in either the header or footer or split information between the header and footer as required 7 Click OK to apply the changes Report items toolbar icons The table below describes the different functions of the report items toolbar icons in the Customize Report window Toolbar icon Function Adds free text Adds a picture xuri Cell Expansion System W25 User Manual 29 0646 22 AA 105 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Toolbar icon Function Adds a chart Includes a method Adds documentation Adds an evaluation log Add objects to the report The table below describes how to add objects to the report Step Action 1 e Click the appropriate icon in the Report items toolbar or e Choose an object from the Insert menu Z e Press and hold the left mouse button on the report page and drag out a box to the size of the item you want to insert Note The mouse pointer shows a symbol for the type of item you have selected e
168. ou want to then find information about select Help Help for in the UNICORN module of interest a UNICORN module noe Tools Contextual Help Fl Instruction Set About UWITORM find information about e press the F1 key with the item of interest selected and the item currently select in focus ed and in focus e g a pane a dialog or a method phase e click the Help icon in the open dialog navigate the online help e select Help Help for in any of the UNICORN modules see illustration above e inthe TOC Table of contents pane expand the headings of interest to navigate the content structure e click the heading of interest to open a section search for a specific e select Help Help for in any of the UNICORN modules term in the online help see illustration above e inthe Search pane enter the term of interest in the input field e click the Search button 38 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 6 UNICORN software overview 2 6 2 The UNICORN help If you want to access manuals in PDF select Help Help for in any of the UNICORN modules format see illustration above in the TOC pane expand the heading UNICORN online documentation portal and select Documentation overview in the PDF manuals section click one of the text links find information about In the Method Editor module n instruction AE UE open a method select
169. ows DO measurement related settings Setting Description Set from View from PP RD Calibration value 100 air Settings DO sensor calibra saturation Cellbag tion values Calibration value tempera Settings DO sensor calibra ture when determining Cellbag tion values clhp Calibration value 0 air Settings DO sensor calibra saturation Cellbag tion values Calibration value tempera Settings DO sensor calibra ture when determining Cellbag tion values clzp calp Calibration value atmo Settings DO sensor calibra spheric pressure during Cellbag tion values calibration Reading The reading cyclewhen DO Settings DO Set DO reading Viewed cycle control is off cycle time when time DO control is off Resetcali A System setting that de System settings DO bration fines if the DO calibration sensor Reset cali valuesat values shall be reset or not bration values at END when the system goes to END END Note In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 277 8 Reference information 8 8 Control settings 8 8 9 DO measurement Output data Description View from PP RD CD DO measurement timer The time left to next DO measurement DO measurement cycle The actual measurement cycle time This value is set
170. p calibration Introduction Calibrating the pumps is in most cases not necessary Setting the tube inner diameter correctly is often enough However if a high precision of the accumulated volume counter is needed calibration is recommended To calibrate a pump 1 click System Calibrate 2 select which pump to calibrate Tube inner diameter Always set the tube inner diameter correctly If the tube inner diameter is not set correctly the following can happen e The wanted flow might appear to be out of range for the given tube diameter That will prevent the media control to start and give warnings in pH control e The pH and media control performance might decline e The pump might not accept a calibration since the calibration result will be too far from the expected Calibration of acid and base pumps The acid and base pumps are calibrated at a fixed speed of 100 rpm That is due to the fact that the working flow of these pumps when using pH control is automatically com puted based on the measured weight and the entered molarity of the acid and base These values might not be representative at the moment of calibration and might also vary during the run Therefore a fixed rom is used for calibration instead of a flow To get a well tuned pH control e Enter the correct tube inner diameter e Enter the correct molarity of acid and base e Make sure that the weight measurement corresponds to the weight of the content in
171. pH and DO control principles 4 1 3 pH and DO control principles Introduction The pH and DO controls have several control modes each The settings for each control mode can be automatic or manual Control schemes and control modes pH control modes Two different pH control modes exist CO2 and Acid Base They are combined to three different control schemes shown in the table below Control scheme Description Control modes used CO2 Base pH control by change of CO concen CO2 tration in the gas mix and by base Acid Base acid addition blocked The software starts with CO2 mode and then switches automatically be tween CO2 and Acid Base mode In Acid Base mode the acid is blocked Acid Base OH control by addition of acid and base Acid Base CO2 pH control by change of CO concentra CO2 tion in the gas mix DO control modes Two different DO control modes exist 02 and Speed They are combined to three different control schemes see table below Control scheme Description Control mode used Speed DO control by change in rocking speed Speed 02 DO control by change of O concentra 02 tion in the gas mix 118 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 1 pH and DO measurement and control 4 1 3 pH and DO control principles Control scheme Description Control mode used 02 Speed e DO control by change of O concen tr
172. page from the report Exit This button closes the Customize Report window Note The general toolbar icons are described below The toolbar icons for specific formatting operations are described in the instructions for how to use the functions General toolbar icons The table below describes the different functions of the general toolbar icons in the Customize Report window Toolbar icon Function Opens a new blank report Note You can also choose the File New menu command Opens the Open Report Format dialog You can choose to open a previously defined format for editing Note You can also choose the File Open menu command Saves the edited report format Note You can also choose the File Save menu command 102 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Toolbar icon Function Cuts the selected object from the report Note You can also choose the Edit Cut menu command Copies the selected object in the report Note You can also choose the Edit Copy menu command Pastes a copied or cut object from the clipboard into the report Note You can also choose the Edit Paste menu command xuri Cell Expansion System W25 User Manual 29 0646 22 AA 103 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 5 Create a new report format Change the page layout The pag
173. pansion System W25 User Manual 29 0646 22 AA 45 3 The UNICORN 3 2 System control Curves 46 software Monitor signals and instrument settings are shown in the chart as curves The curves are also saved in a result file which can be opened in the Evaluation module The default view shows the most commonly used curves The user may customize which curves to display and the color and style of the displayed curves Note Note All curves are saved in the result file regardless of the curves displayed The complete ranges of the signals are saved in the result file regardless of used factors such as scaling and zooming during the run The table below describes how to customize which curves to show in the chart Refer to the online help for further information about the tabs in the Customize dialog Step Action 1 In the System Control module click the Customize icon i or select Tools Customize Result The Customize dialog opens Select the Curves tab Customize System 2 Run Data Groups il Curve Style and Color XAxis Y Axis Run Log Select curves to display 001 Rocking speed C 002 Rocking angle setpoint V 003 Weight V 004 Temperature C 005 Temperature setpoint V 006 Gasflow C 007 Gasflow setpoint F 008 c02 C 009 CO2 setpoint i 010 02 _ 013 02 setpoint V 014 pH C 015 pH setpoint C O16 pH amplitude C 018 pH phase O 019 pH acid
174. pansion W25 CBCU and Cellbag bioreactor Tygon E3603 Tubing between gas distribution tubing and the inlet filter of the Cellbag bioreactor Silicone VMQ Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 5 Chemical resistance The chemicals listed below have been approved for use with the bioreactor system Chemical Concentration Alconox Cleaning Disinfection 8 Reference information 8 5 Chemical resistance CAS no EC no N A DeciDos Cleaning Disinfection N A Ethanol Cleaning Disinfection 75 08 1 200 837 3 Hydrochloric acid pH control 7647 01 0 231 595 7 lsopropanol Cleaning Disinfection 67 63 0 200 661 7 Klercide Cleaning Disinfection N A PBS solution Testing N A Sodium bicarbonate OH control 144 55 8 205 633 8 Sodium carbonate OH control 497 19 8 207 838 8 Sodium chloride Sodium hydroxide Testing OH control 7647 14 5 231 598 3 1310 73 2 215 185 5 Sodium hypochlorite Cleaning Disinfection 7681 52 9 231 668 3 Virkon Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Cleaning Disinfection N A 245 8 Reference information 8 6 Tubing and connectors 8 6 Tubing and connectors The tubing and connectors listed below are delivered with the bioreactor system The user cuts the appropriate lengths for the tubing and connects all connectors needed for the appl
175. plicable 264 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 7 pH measurement 8 8 7 pH measurement pHOPT calibration values The pH surrounding the pH sensor is calculated from a measured variable called the phase angle The calculation is based on a transfer equation programmed in the pHOPT monitor This transfer equation is defined by a set of constants or more correctly a set of values that are specific for the mixture of the substances forming the luminophoric dye This means that the dye properties and consequently the transfer equation may differ between different dye batches The constants for each batch after gamma irradiation are determined at the factory and are printed on the pHOPT label of the Cellbag as calibration values cmin cmax cpHO and cdpH The interpretation of the calibration values on the transfer equation is illustrated in the graph below Phase angle cmin _point of inflexion mn Cmax pH epHo A fifth calibration value ctemp is also necessary It is not directly included in the transfer equation but since the sensor is temperature dependent the ctemp value is used for fine tuning the pH against the actual temperature xuri Cell Expansion System W25 User Manual 29 0646 22 AA 265 8 Reference information 8 8 Control settings 8 8 7 pH measurement Settings Setting The following table shows pH meas
176. point allowed for the pH control tion Min CO2 The minimum CO concentration pH control CO2 Y concentra setpoint allowed for the pH control tion PID parame When Auto is selected the system pH control A hand ter mode will automatically set the PID param advanced CO2 symbol is eters When Manual is selected the shown if system will use the manual PID pa set to rameters set by this instruction Manual Manual P The P parameter used if PID param pH control Y eter mode is set to Manual advanced CO2 Manual I The l parameter used if PID param pH control Y eter mode is set to Manual Note advanced CO2 that increasing the l parameter will reduce the integrating effect and vice versa xuri Cell Expansion System W25 User Manual 29 0646 22 AA 269 8 Reference information 8 8 Control settings 8 8 8 pH control Setting Description Set from View from MI PP RD CD Manual D The D parameter used if PID param pH control ad eter mode is set to Manual vanced CO2 Cycle time When Auto is selected the system pH control ad A hand mode will automatically set the cycle time vanced CO2 symbol is When Manual is selected the sys shown if tem will use Manual cycle time set to Manual Manual cy The cycle time used if Cycle time pH control ad cle time mode is set to Manual vanced CO2 Note In the table the following abbreviations are used PP Process P
177. pply pressure is between 1 0 and 1 5 bar Erratic O control pH control Error symptom O supply is not proper ly connected Check that the O gas is properly connected and at the correct pressure 1 0 to 1 5 bar Incorrect setpoint Check the setpoint Incorrect gas supply air or N3 is connected Check that the gas supply chosen in UNICORN matches the connected gas supply air or N3 O sensor is broken Possible cause Contact GE Healthcare service personnel Corrective action pH control is not start ing Some of the criteria for starting the pH control are not fulfilled Read the message in the pop up dialog If possible take action according to the information and try again 214 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Error symptom PH control using CO seems to react too slow or too fast undershoot ing overshooting Possible cause The bag size setting is incorrect which will op timize the control for another gas volume The control is using manually entered con trol parameters not op timal for the actual case 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Corrective action Set the bag size to the correct value Use the manual instruction pH control ad vanced pH control advanced CO2 to set PID parameter mode and Cycle time mode to Auto See Manual instructions on page 47 The CO range is too
178. pump can be performed In the pH control A B C case it should however be enough just or D to give the tubing diameter PID When Auto is selected the system will OH control A parameter automatically set the PID parameters advanced hand mode When Manual is selected the system acid base sym will use the manual PID parameters set bol is by this instruction shown if set to Manu al Manual P The P parameter used if PID parameter OH control Y mode is set to Manual advanced acid base Manual I The l parameter used if PID parameter pH control Y mode is set to Manual advanced acid base Cycle time When Auto is selected the system will pH control A mode automatically set the cycle time When advanced hand Manual is selected the system will use acid base sym Manual cycle time bol is shown if set to Manu al Manualcy The cycle time used if Cycle time mode pH control Yy cle time is set to Manual advanced acid base 272 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 8 pH control Setting Description Set from View from MI PP RD CD Flow mode When Auto is selected the system will automatically set the acid base flow The automatically computed acid base can be displayed as run data When Manual is selected the system will use the manual acid base flow set by this instruction pH control advanced acid base
179. put in percent This run data is not specifically connected to the feed or harvest role Instead this run data is connected to a specific pump head Look at the run data from the pump head of interest Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable xuri Cell Expansion System W25 User Manual 29 0646 22 AA 251 8 Reference information 8 8 Control settings 8 8 2 Heating control 8 8 2 Heating control Heating control settings Setting Temperature setpoint Description The temperature setpoint of the liquid in the Cellbag Settings Temp The following table shows general settings used for heating control Set from Heating View from PP RD Deviation alarm Select On to activate the devia tion alarm and Off to deactivate it Heating Allowed devia tion up A parameter that defines how much above the setpoint the temperature reading is allowed to be without triggering the de viation alarm Settings Temp Heating Allowed devia tion down A parameter that defines how much below the setpoint the temperature reading is allowed to be without triggering the de viation alarm Settings Temp Heating Deviation alarm delay A parameter that defines for how long the temperature read ing can be outside the allowed deviation limits before the
180. r that you want to calibrate Result The Calibration dialog opens E x v Current value 5 221 kg Calibration P2 4 07 Monitor to calibrate Weight Calibration procedure Enter reference weight low point 1 Ensure the rocker is placed on a stable leveled 0 50 and even surface gt 2 Set the rocker stop angle to 0 deg Calibrate low point 0 0000 kg Calibrate low point 3 Start the rocking then stop the rocking and 0 50 lensure the rocker is stopped with the tray in horizontal position 4 Under Settings Weight in the process picture _ ensure that the weight distribution between the four feet is even by checking that the numbers are all 25 5 Adjust the right front foot if needed 5 Enter the low point weight as 0 0 and press Calibrate low point 6 Put a well defined weight mass gt 10 kg uncertainty lt 0 001 kg centred on tray Enter the known weight and press Calibrate high point Calibrate High point 10 0000 kg Last calibrated on DEO OT PATT ETI Se E Pon Select the appropriate monitor in the Monitor to calibrate drop down menu Follow the instructions in the right hand field and enter the correct values in the Calibration procedure field and click Calibrate for each value Close the Calibration dialog 202 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 6 Maintenance 6 2 Pump calibration 6 2 Pum
181. r panel Reposition the connectors or insert jumpers in the unused positions between the occupied ports See Chapter 4 Installation in the Operating Instructions Two pumps are con nected but only one pump is defined in UNICORN Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Read the displayed message when you connect to the system in System Control Make sure all available components are selected for the system See Section 5 3 Edit system properties on page 174 207 7 Troubleshooting 7 2 Xuri Cell Expansion System W25 rocker 7 2 Xuri Cell Expansion System W25 rocker Rocking Error symptom Rocker does not initial ize properly Possible cause Faulty safety fuses Corrective action Try to restart the system a few times If the system still does not initialize properly shut it down and contact GE Healthcare service person nel When starting the run the rocker moves too slowly during the first cycle It takes a while for the rocker to initialize after turning on the power None This is normal Rocker is not rocking Rocker is in an error state e Check the current alarms e Reset power to the unit e lf you still get motor alarms contact GE Healthcare service personnel Tray is in tilt position e Make sure that the tray is in normal position e lf this does not help contact GE Healthcare service personnel Rocker stops rocking
182. re 3 4 Evaluation in UNICORN 3 4 1 Evaluation 3 4 1 Evaluation Introduction The Evaluation module is used to evaluate the results from bioreactor runs Evaluation panes As illustrated below the Evaluation module contains two panes When a result is opened from the Result Navigator 1 the Chart pane 2 is displayed In the Evaluation module it is also possible to view the complete documentation of the results and to generate reports Refer to the UNICORN Online Help for more information about result evaluation amp Evaluation AA perf EJ File gdt View Tools Help Jael 7 Weight_Chart 1 44 pert Temperature_Chart 1 44 perf 02_Chart 1 4A perf DO_Chart 1 AA perf Folder name A CO2_Chart 1 44 perf f HCE GDBBN4 amp DefaultHome amp Bioreactor Lol AA pert all pH 3 to 12 1 fdbpH test 2 to 12 a pH test 3 to 12 z fx Procedure B system Evaluation toolbar icons The table below shows the Evaluation toolbar icons that are referred to in this User Manual Open Result Navigator Opens the Result Navigator where available re sults are listed Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 87 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 1 Evaluation Function Save Saves the changes made to the current result Print Opens the Print charts dialog from where a chart can be printed Report Opens the Create repor
183. reactor gel 7 6 Software problems 229 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 7 Troubleshooting 7 1 Xuri Cell Expansion System W25 7 1 Xuri Cell Expansion System W25 Alarm messages For many of the problems that may occur UNICORN displays an alarm message on the screen Follow the instruction shown to solve the problem System units not recognized Error symptom CBCU not recognized by UNICORN Possible cause Incorrect CBCU CAN ID Corrective action Check and if necessary change CAN ID using the switch on the CBCU rear panel See CAN ID on page 28 The CBCU CAN ID should always be set to position 1 The system must be switched off before the CAN ID is changed and then restarted CBCU is not selected as a component Click Edit in System Properties in the Administra tion module and add the CBCU as a component Pump not recognized by UNICORN Incorrect pump CAN ID Check and if necessary change CAN ID using the switch on the pump rear panel See CAN ID on page 31 The pump CAN ID should be set to posi tion 1 or 2 and the pumps must have unique val ues The system must be switched off before the CAN ID is changed and then restarted Pump Is not selected as a component Click Edit in System Properties in the Administra tion module and add the pump as a component CBCU and or pump not recognized by UNICORN Gap between occupied UniNet 9 ports on the rocker rea
184. ressing the power button to force system shut down If problem persists contact GE Healthcare service personnel Rocking starts when sending another com mand than Start rock ing At the first command sent to the system the system enters Run state and the rocker is initialized and makes a few movements Xuri Cell Expansion System W25 User Manual 29 0646 22 AA None This is normal 209 7 Troubleshooting 7 2 Xuri Cell Expansion System W25 rocker Temperature Error symptom Possible cause Corrective action No heating although Make sure that the rocker is rocking The heater is heating is enabled in automatically turned off when rocking is turned UNICORN Frame off around the tempera ture button is orange Rocker is not rocking The recognition of the tray size used does not work Contact GE Healthcare service personnel Bag size is not set Set the bag size in Settings Cellbag in the Process Picture The heater is in an error state Contact GE Healthcare service personnel Too slow or too fast heating Bag size setting Is incor rect Check and if necessary reset the bag size setting When changing the bag size the rocking must be turned off Incorrect weight mea surement Make sure that the rocker is placed in a hori zontal position Check that the weight shown in the Process Picture matches the actual weight of the cul ture medium If not tare t
185. rmed by a rocker unit The system composed of the rocker CBCU and pump enables measurement and control of pH DO weight and media distribution and provides different gas flow and gas mixing possibilities Illustration of the system The illustration below shows the main bioreactor system units 1 2 5 4 mmmn 8 7 6 5 Part Description Hatch 2 Filter heater 3 Cellbag bioreactor xuri Cell Expansion System W25 User Manual 29 0646 22 AA 13 2 System description 2 1 System overview Description Xuri Cell Expansion W25 Pump Xuri Cell Expansion W25 CBCU xuri Cell Expansion System W25 rocker Tray Lid Illustration of wave motion Description Inflowing gas from the CBCU enters the Cellbag bioreactor through the inlet gas filter The gas flow inflates the bag and oxygenates the culture Metabolic waste gases leave the Cellbag bioreactor through the outlet gas filter The pressure control valve maintains a constant overpressure inside the Cellbag bioreactor The rocking mechanism sets the rocking platform in motion Wave motions are induced by the rocking The culture is cautiously mixed and gases are transferred into the culture 14 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 1 System overview Available control parameters The table below describes the control parameters for the fully configured bioreactor system
186. rom the expected 204 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 6 Maintenance 6 3 Cleaning 6 3 Cleaning Cleaning procedure To prevent microbial or cross contamination Xuri Cell Expansion System W25 should be cleaned after each cultivation The system must be turned off and unplugged before cleaning begins e Clean the exterior of the system units with a damp cloth and a suitable cleaning agent e Make sure to clean the temperature sensor arms on the underside of the rocker platform If dirt accumulates in these arms they may not function properly which could cause incorrect temperature regulation Recommended cleaning agents All system units can be cleaned with ethanol isopropanol and Virkon with suitable concentrations Refer to Section 8 5 Chemical resistance on page 245 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 205 7 Troubleshooting 7 Troubleshooting In this chapter This chapter describes troubleshooting and corrective actions for Xuri Cell Expansion System W25 If the suggested actions in this guide do not solve the problem or if the problem is not covered by this guide contact your GE Healthcare representative for advice This chapter contains the following sections Section See page 7 1 Xuri Cell Expansion System W25 207 7 2 Xuri Cell Expansion System W25 rocker 208 7 3 Xuri Cell Expansion W25 CBCU 212 7 4 Xuri Cell Expansion W25 Pump 226 7 5 Cellbag bio
187. roperties dialog Note Only active systems can be edited e Click the Edit button Result The Edit dialog is displayed Edit System 2 Instrument configuration WAVE 25 1 0 0 0 vi Instrument server EPC 123456 Instrument serial no 123456 Connect by Fixed IP address f Instrument serial no Rare ras Component types Selections Component selection Property CBCU Pump 25 1 Pump 25 2 3 1 Select the desired instrument configuration from the Instrument config uration droplist menu 2 Select the instrument server named EPC lt serial number gt from the In strument server droplist menu 3 Choose the connection type Instrument serial no 4 All possible components are shown in the Component selection list e Click the checkboxes to select or de select components e Make sure the components selected match the units connected to the system 9 Click the OK button to apply the changes xuri Cell Expansion System W25 User Manual 29 0646 22 AA 175 5 Operation 5 4 Prepare cultivation 5 4 Prepare cultivation In this section 176 This section describes the procedures before starting the cell cultivation For illustrations and descriptions of the system refer to Chapter 2 System description on page 12 This section contains the following subsections Section See page 5 4 1 Start a run 177 5 4 2 Pre culture preparation 182 5 4 3 Final checks before cultivation 184 5 4 4 Ad
188. roximately 1 air saturation 10 mbar xuri Cell Expansion System W25 User Manual 29 0646 22 AA 115 4 System control description 4 1 pH and DO measurement and control 4 1 1 pH and DO measurement principles At delivery the DO configured Cellbag bioreactors have sensors that are factory calibrated at a certain atmospheric pressure This pressure is stated on the DO label as Calp and shall be entered together with the other calibration values for DO To compensate for differences in atmospheric pressure from where the bag was calibrated to where the cell culture will be performed it is possible to re calibrate the sensor To do this select System Calibrate DO sensor in UNICORN O measurement and altitude 116 The O sensor measures the relative O concentration in terms of vol The O concen tration in the atmosphere Is 21 vol independently of altitude above sea level and weather conditions Since the most relevant parameter from a cell cultivation perspective is the dissolved oxygen expressed as mg l different cell growth conditions will be obtained depending on the ambient pressure For instance at 2000 m above sea level the ambient pressure is 0 8 bar 800 hPa The relative O concentration is 21 vol but compared to the conditions at sea level where the ambient pressure is 1 013 bar 1013 hPa the absolute O concentration in ambient air at 2000 m corresponds to sini 16 6 vol 1013 0 at sea level Different growth con
189. rt Protocol Start Protocol ri Scouting or e click the Method Settings phase and click the Start Protocol button in the Phase Properties tab Start Protocol Result The Start Protocol dialog opens 2 In the Start Protocol dialog e Select items to display at method start When selecting a method item a description is shown to the right Result Name and Location is selected by default e Click OK to confirm and close the dialog Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 61 3 The UNICORN software 3 3 Methods in UNICORN 3 3 3 Work with methods Add edit Method Notes The table below describes how to add edit notes to a method Step Action 1 In the Method Editor e click the Method Notes icon or Ctrl z Result Name amp Location Method Nokes i e click the Method Settings phase and click the Method Notes button in the Phase Properties tab Method Notes Result The Method Notes dialog opens 62 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 3 Work with methods Step Action 2 Method Notes In the Method Notes dialog e Enter edit notes about the method If notes already have been entered it is possible to search for specific words using the Find button e Click OK to confirm and close the dialog Xuri Cell Expansion System W25 User Manual 2
190. ructions so any functional instructions must be added by hand To include instructions for the User Defined phase select the Text Instructions tab The Phase Properties tab will only show the variables used in this phase See Section 3 3 4 Text in structions on page 64 for information about how to work with instructions in the Text Instructions pane xuri Cell Expansion System W25 User Manual 29 0646 22 AA 59 3 The UNICORN software 3 3 Methods in UNICORN 3 3 3 Work with methods Rearrange phases within a The table below describes how to rearrange phases within a method Action Select the phase to be moved in the Method Outline pane e Drag and drop the phase to the requested position in the Method Outline pane Result The phase is moved to the requested position Or e Right click the phase and select Move up or Move down User Defined Rename Delete Save Phase User Defined ave Pease Move Down Cut Copy Result The phase is moved one step up or down in the Method Outline method Step 2 60 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 3 Methods in UNICORN 3 3 3 Work with methods Set up a Start Protocol The table below describes how to set up a Start Protocol to be displayed before the method run starts Step Action 1 In the Method Editor e click the Start Protocol icon mm T E e select Tools Sta
191. rument Ethernet Power supply 100 120 200 240 V 50 60 Hz Power consumption Maximum 1500 VA Enclosure protective class IP 21 External air supply 1 0 bar to 1 5 bar Normal use 1 3 min Fast fill 3 5 l min External CO supply 1 0 bar to 1 5 bar Normal use 0 2 min Fast fill 0 5 I min External O supply 1 0 bar to 1 5 bar Normal use 0 7 min Fast fill 1 7 I min Operating ambient temperature range 15 C to 32 C Operating humidity range Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 20 to 80 RH non condensing 237 8 Reference information 8 2 Component specifications 8 2 Component specifications Equipment dimensions The table below lists the outer dimensions of the bioreactor system units System unit Rocker Dimensions W x D x H mm 404 x 560 x 205 CBCU Pump 276x 360x117 275 x 280 x 115 Tray 10 475 x 430 x 60 Tray 20 740 x 480 x 70 Tray 50 800 x 610 x 70 Lid 10 475 x 430 x 230 Lid 20 740 x 480 x 245 Lid 50 Equipment weight 800 x 610 x 260 The table below lists the weights of the bioreactor system units System unit Rocker Weight kg 24 0 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Rocker specifications 8 Reference information 8 2 Component specifications The table below he table below lists the s
192. s below Turn off the DO reading until the culture medium is equilibrated DO fiber optical cable is not connected properly or is defective Check that the correct calibration values as stated on the Cellbag have been entered See Sec tion 5 4 4 Addition of culture medium on page 185 for more information Note The value Clhp and Clht may have changed after recalibration The initial DO reading is much higher than ex pected approximately 300 air saturation when 100 air satura tion is expected DO fiber optical cable is not connected properly or is defective Check both ends of the fiber cable Make sure that all four pins of the sensor port is gripping the sen sor adapter and that the fiber cable is properly connected to the DO port on the CBCU front panel If both pH and DO sensors are used check that the fiber cables are not mixed up If so correct the connections of the pH and DO sensors Note that it may take one reading cycle before correct values are shown Check that the correct calibration values as stated on the Cellbag have been entered See Sec tion 5 4 4 Addition of culture medium on page 185 for more information Note The value Clhp and Clht may have changed after recalibration 222 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Error symptom The initial DO reading deviates from what is expected error is typi cally around 10 air saturat
193. s xP Windows 7 Intel Dual Core or better Windows XP Windows 7 Windows Server 2003 x86 x64 R2 x86 X64 Windows Server 2008 x64 R2 x64 Intel Dual Core or better Windows XP Windows 7 Windows Server 2003 x86 x64 R2 x86 X64 Windows Server 2008 x64 R2 x64 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 3 Client computer specifications Note e UNICORN is tested using an English operating system version Using other language versions of the operating system may cause errors e A screen resolution of 1280x1024 or higher is recommended Parts of the UNICORN user interface may not be displayed properly using a lower res olution e Changing the default font and font size in Windows may cause problems in the UNICORN user interface e The Windows basic color scheme is recommended e Using the Windows 7 Aero color scheme is not recommended e Windows power save features should be turned off to avoid conflicts with system operations e UNICORN is not compatible with the Windows 7 feature High DPI Aware ness which allows the graphic user interface to be scaled The interface scale must remain at 100 to avoid issues with clipping and misaligning of parts of the UNICORN user interface Normally the scale is set at 100 by default 1 UNICORN must be closed when the color scheme is changed Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 243 8 Reference i
194. s below 21 to lower the DO although N gt is connected to CBCU The setting for gas source is not set to N2 Select N as gas source so that the DO control can command O setpoints below 21 220 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Error symptom DO control using speed seems to react too slow or too fast undershoot ing overshooting Possible cause The entered parame ters for cycle time speed step max min speed and deadband are not optimal for the actual case A gas bubble might be present on the DO sen Sor 7 Troubleshooting 7 3 Xuri Cell Expansion W25 CBCU Corrective action Laborate with the parameters There is no auto matical parameter mode for DO control using speed Tap the DO sensor from the underside to remove the bubble This may require forceful manipulation and the Cellbag bioreactor may need to be partly disconnected from the tray Note The gas bubble may be very difficult to see In control scheme 02 Speed the transi tions between O and speed are longer or shorter than desired The automatically computed transition delay time is not opti mal for the actual cell culture Check what the actual transition delay time values are by selecting them as run data under Tools Customize Use the manual instruction DO control advanced DO control transition delay Set O2 Speed transition delay mode to Manual and enter desired values for trans
195. s the light signals Light and temperature sensitivity The sensor is subject to photobleaching which means its properties are affected by light This includes ambient light as well as the light from the LED To reduce the photobleaching make sure not to unnecessarily expose the Cellbag to light For example e Keep the Cellbag in its protective black bag until shortly before use e Use the Rocker lid The LED output is automatically regulated to provide suitable amplitude for optimal performance and life time of the sensor The sensor is also temperature sensitive The pH and DO modules include temperature compensation for use within a few C around the calibration temperature DO air saturation and atmospheric pressure DO is measured as percent air saturation 100 air saturation represents the oxygen content of a certain solution at the current temperature and atmospheric pressure when it has been equilibrated in an environment of air Air saturation is a relative measurement The oxygen content measured in for example mg l depends on factors such as the composition of the solution temperature and atmospheric pressure due to different weather conditions or altitude If a DO sensor is calibrated for 100 air saturation under equilibrated conditions at one atmospheric pressure such as 950 mbar it will not show 100 air if the same content is later equilibrated at another atmospheric pressure say 1050 mbar The difference is app
196. scheme Save the method xuri Cell Expansion System W25 User Manual 29 0646 22 AA 81 3 The UNICORN software 3 3 Methods in UNICORN 3 3 7 Method queues 3 3 7 Method queues Introduction This section describes how to create and edit method queues in UNICORN For information on how to create and edit individual methods see Section 3 3 2 Method creation on page 53 A method queue in UNICORN is a linked set of methods to be run The method queue can contain methods to be run on up to three different systems Each system may have up to ten methods queued Create a method queue 82 The following table describes how to create a method queue Step Action 1 In the Method Editor e click the New Method Queue icon in the Toolbar or e Select File New Method Queue Ji wiew Phases Tools J New Method Ctrl M New Method Queue Result The Method Queue dialog opens Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Step 3 The UNICORN software 3 3 Methods in UNICORN 3 3 7 Method queues Action In the Method Queue dialog choose the Number of included systems from the drop down list Method Queue Number of included systems 1 Start Condition v Result A separate method queue block will be added to the dialog for each additional system if required Choose a system for each method queue block from the System drop down list System P2 9 01 w Method m
197. sed without editing after the end of the method run Variable List The Variable List tab shows all the method variables and corresponding values listed by the method block where they appear The list can show detail and or unused variables At this stage the variable values are no longer possible to edit xuri Cell Expansion System W25 User Manual 29 0646 22 AA 95 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 3 Run documentation Documentation tab Contents Scouting The Scouting tab shows the scouting parameter settings This tab corresponds to the Scouting settings in the Method Editor Text Instructions The Text Instructions tab shows all the text method in structions from the method These instructions can be saved as a new method This is described in Save the method used for the run as a new method on page 97 in this section The Notes tab shows all notes that have been made re garding the method the method run and the subsequent evaluation The notes are divided into the following sub tabs e Method Notes Start Notes Run Notes e Evaluation Notes You can add new notes on the Evaluation Notes sub tab Note Click the Find button to search for specific text in the Notes Search for log entries The table below describes how to find specific text in the logs Step Action 1 Select the log tab where you want to perform your search 2 e Click the Find button Result The Fin
198. ser Manual 29 0646 22 AA 275 8 Reference information 8 8 Control settings 8 8 9 DO measurement 8 8 9 DO measurement DOOPT calibration values 2 6 The DO surrounding the DO sensor is calculated from a measured variable called the phase angle The calculation is based on a transfer equation programmed in the DOOPT monitor This transfer equation is defined by a set of constants or more correctly a set of values that are specific for the mixture of the substances forming the luminophoric dye This means that the dye properties and consequently the transfer equation may differ between different dye batches The constants for each batch after gamma Irradiation are determined at the factory and are printed on the DOOPT label of the Cellbag as calibration values clhp and clzp The interpretation of the calibration values on the transfer equation is illustrated in the graph below Phase angle clzp cihp DO 100 air saturation The calibration values clht and clzt are temperatures recorded during the factory cali bration They are used by the DO monitor for temperature compensation The calibration value calp is the atmospheric pressure during the factory calibration It is not used by the DO monitor but is kept for informative purpose Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 9 DO measurement Settings The following table sh
199. sert button Result The instruction will be inserted in the block e atthe position of the breakpoint of the new instruction if there are no other instructions at that breakpoint e immediately after the currently highlighted instruction if the highlight is at the same breakpoint as the new instruction e as the first or last instruction if none of the instructions at the desired breakpoint is highlighted The insertion point depends on the breakpoint value of the currently selected instruction E g if current instructions breakpoint is lower than the breakpoint value for the new instruction it is inserted as the first instruction at that breakpoint xuri Cell Expansion System W25 User Manual 29 0646 22 AA 65 3 The UNICORN software 3 3 Methods in UNICORN 3 3 4 Text instructions Define new variables 66 Only one variable that affects block length breakpoint may be defined within each block However any number of parameters may be defined as variables within a block The table below describes how to define a new variable Step Action 1 Select the instruction where you want to define the variable in the Text In structions area Result The parameters for the instruction are shown in the Instruction Box 2 e Locate the breakpoint or the required parameter in the Instruction box e Click the Var button 0 00 Base SameAsMain 0 00 Rocking 20 0 Rocking Speed RPM 6 0 Rocking Angle o 12 0 o 5 min
200. set so that sufficient mixing and oxygen transfer is provided without excessive foaming e The culture volume affects mixing and oxygen transfer rate Lower volumes give faster oxygen transfer and mixing e Increasing the rocking speed and angle will result in faster oxygen transfer and mixing e f no clear wave appears inside the Cellbag bioreactor change the rocking speed most often an increase is needed e For cultures with high oxygen demand such as a suspension CHO cells rocking speed and angle can be increased up to for example 30 rom and 8 degrees respec tively e For shear sensitive cells such as adherent cells on microcarriers rocking speed and or angle can be decreased down to 10 rpm and 4 degrees if sufficient oxygen transfer and mixing is maintained Each new cell line and process requires optimization of operating conditions The table below shows typical rocking speed and gas flow settings for suspension cell culture Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 141 4 System control description 4 5 Recommended operating conditions Cellbag biore Culture vol Rocking Rockingangle Gas flow lpm actor size L ume L speed rpm 2 0 3 10 20 2 4 0 05 0 2 Note When cultivating ina 50 L Cellbag bioreactor at maximum working volume of 25 L rocking speed and angle multiplied should not exceed 240 rom degrees For example if the rocking angle is set to 8 degrees
201. signal sounds and End is displayed in the Run Log This also applies when ending a manual run Tip In System Settings it is possible to set Prepare for Tilt at END If you want to temporarily hold the method click the Hold icon Q Note When a method is held the system control is maintained but no new instructions are given resume a held method run click the Continue icon Ud Note An ended method cannot be continued resume a method run ora manual click the Continue icon run after the system has entered the Error and Alarm state lb permanently end the run click the End icon Note When ending a method run prematurely it is possible to save the partial result 180 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 4 Prepare cultivation 54 1 Starra run Actions in the Process Picture pane It is possible to interact with the Process Picture Refer to Actions in the Process Picture pane on page 43 for more information xuri Cell Expansion System W25 User Manual 29 0646 22 AA 181 5 Operation 5 4 Prepare cultivation 5 4 2 Pre culture preparation 5 4 2 Pre culture preparation Instructions Follow the instructions below to prepare the bioreactor system for cell cultivation NOTICE Always use aseptic technique when connecting or disconnecting tubing to the Cellbag bioreactor Step Action 1 Follow these instructions to inflate
202. stem W25 User Manual 29 0646 22 AA 15 2 System description 2 2 Xuri Cell Expansion System W25 rocker 2 2 uri Cell Expansion System W25 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 an embedded computer which allows running the system independently of the performance of the connected network and client computer The dimensions and weight of the rocker are listed in Section 8 2 Component specifications on page 238 Rocking parameters 16 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 re sulting in a step wise rocking The maximum value 100 gives a sinusoidal motion with Slower angular velocity in the end positions and faster in the middle of the movement resulting in a smoother rocking Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 2 Xuri Cell Expansion System W25 rocker Front view of
203. stration and Technical manual Overview and detailed description of network setup and complete software installation Administration of UNICORN and the UNICORN database UNICORN Online Help Dialog descriptions for UNICORN User Documentation CD Xuri Cell Expansion System W25 User Manual 29 0646 22 AA CD containing the listed manuals and translated versions of Xuri Cell Expansion System W25 Operating Instructions 11 2 System description 2 System description In this chapter 12 This chapter gives an overview of Xuri Cell Expansion System W25 and describes the different bioreactor system units This chapter contains the following sections Section See page 2 1 System overview 13 2 2 Xuri Cell Expansion System W25 rocker 16 2 3 Xuri Cell Expansion W25 CBCU 26 2 4 Xuri Cell Expansion W25 Pump 29 2 5 Cellbag bioreactor 32 2 6 UNICORN software overview 36 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 1 System overview 2 1 System overview Introduction Xuri Cell Expansion System W25 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 The cell culture volume range is 0 3 L to 25 L and working volume may be expanded up to 10 times during one cultivation Gas transfer and mixing of culture is accomplished by wave induced agitation perfo
204. t of the cell culture medium See Section 5 4 4 Addi tion of culture medium on page 185 for more infor mation The control is using manually entered con trol parameters not op timal for the actual case Use the manual instruction pH control ad vanced pH control advanced acid base to set PID parameter cycle time and flow mode to Auto See Manual instructions on page 47 The automatically se lected control parame ters are not optimal for the actual case Use the manual instruction pH control ad vanced pH control advanced acid base Set PID parameter mode Cycle time mode and or Flow mode to Manual and enter desired values for PID parameters cycle time and flow See Manual in structions on page 47 To get an initial suggestion of the PID parameters look at the automatically selected parameters by selecting them as run data under Tools Customize OH control using acid or base is doing too many small injections instead of doing larger ones more seldom PH control using acid or base is doing large injections with long time distance instead of doing smaller ones more often The deadband is set too low The deadband is set too high Check what the actual deadband values are by selecting them as run data under Tools Customize Use the manual instruction pH control ad vanced pH control advanced acid base n crease the deadband value for acid and or base See Manual instru
205. t dialog where a report of the result can be created View Documentation Opens the Documentation dialog that contains the complete documentation for a manual or a method run Customize Opens the Customize dialog where for example curve settings can be set 88 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 2 Open and view results 3 4 2 Open and view results Introduction All contents of the result files are opened in the Evaluation module where you can analyze the results and compile reports The Evaluation module user interface and toolbar icons are described in Section 3 4 1 Evaluation on page 87 This section also describes how to highlight curves in a chart read curve values using a marker and save curve data as a Snapshot Open a result in the Evaluation module Click the Open Result Navigator icon to access the result files that are located in folders accessible to you There are four ways to open a result from the Result Navigator e Select a result and click the Enter key or e Double click a result or e Right click a result and choose Open from the shortcut menu or e Select a result and click the Open toolbar icon in the Result Navigator Open Result The result is opened in the chart pane Only one result at a time may be opened this way If you open a new result the previous result will automatically close However you may ope
206. t to activate or deactivatea click on the right hand side of the button function Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 43 3 The UNICORN software 3 2 System control If you want to open the settings for a function adjust the settings 44 click on the left hand side of the button The example below shows the settings for dissolved oxy gen DO Settings Do Reading Cycle time 5 1000s Control Setpoint OK pH Temp Media control Gas control Cellbag pumps Cellbag Weight Rocking enter appropriate values in the Settings dialog and click OK or press enter Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 2 System control System Control toolbar icons The table below shows the System Control toolbar icons that are referred to in this User Manual Function Open Method Navigator Opens the Method Navigator where available methods are listed Run Starts a method run Hold Suspends the method run Continue Resumes for example a held method run End Permanently ends the method run Customize Opens the Customize dialog where curve settings run data groups and run log contents can be set Connect to Systems Opens the Connect to Systems dialog where systems can be connected and currently connected users are displayed xuri Cell Ex
207. ted in the center of a sensor port on the Cellbag bioreactor and must be coupled to a sensor adapter see table below Description Bag sensor The sensor port is located on the underside of the Cellbag bioreactor port The actual sensor white for pH pink black for DO is located in the center 1 of the sensor port see image below The sensor adapter is attached to the sensor port by the four pins 2 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 2 System description 2 5 Cellbag bioreactor Description Sensor The sensor adapter is located at one end of an optical fiber cable adapter The optical lens of the fiber cable is located in the center of the sensor adapter The fiber cable is connected to a sensor reader in the CBCU The fiber cable is connected to the pH or DO port on the CBCU front panel xuri Cell Expansion System W25 User Manual 29 0646 22 AA 35 2 System description 2 6 UNICORN software overview 2 0 UNICORN software overview In this section 36 This section gives an overview of the general operation of the UNICORN software a complete package for control supervision and evaluation of cell cultivation runs It also describes how to access the help utility that is included in UNICORN Refer to Chapter 3 The UNICORN software on page 40 for more information This section contains the following subsections Section See page 2 6 1 General UNICORN operation 37 2 6 2 The UN
208. ter a while the setpoint is decreased Weight Weight setpoint A _ Weight measurement Cc oO par i wv gt Cc oO aR Feed only Time Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 4 Media control 4 4 2 Perfusion Pump calibration In perfusion mode the true feed flow is dependent of how well the feed pump is calibrat ed When auto calibration is not used at least the feed pump must be calibrated manually It is however recommended to also calibrate the harvest waste pump since unnecessary warnings for harvest waste filter clogging might be given otherwise The accumulated volume will be more precisely computed if the pump is calibrated For a detailed description of pump calibration see Section 6 2 Pump calibration on page 203 Auto calibration 136 Auto calibration is a convenient and precise way of letting the system calibrate the feed and harvest waste pumps automatically The system runs the feed and harvest waste pumps one at the time carefully measures the weight before and after running the pump and then computes the weight difference used as calibration input to the pump See Section 8 7 Auto calibration on page 247 for more information about the auto calibra tion procedure A great benefit with the auto calibration is that it is done on the complete setup No factors will differ between the calibration and run No open ends of tu
209. the Cellbag bioreactor 1 Make sure that all ports on the Cellbag bioreactor are closed and that inlet and outlet filters are open 2 Open Settings Gas control Gas flow from the Process Picture in System Control 3 Enable Fast fill o2 Setpoint 1 00 l min co2 Alert O0 05 0 05 Tania Fast fill Enable Disable Cellbag pumps OK Cancel Cellbag Weight Rocking 4 Turn on Gas flow from the Process Picture by pressing the right hand side of the Gas flow button Note If Fast fill is enabled the gas flow is maximized during the first 20 minutes Result The Cellbag is inflated 182 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 4 Prepare cultivation 5 4 2 Pre culture preparation Step Action 2 Connect inlet and outlet tubing to the Cellbag bioreactor for example acid base feed and harvest waste 3 Adjust the pump parameters for each pump under Settings Cellbag pumps Enter the Tube inner diameter and if the pump function is acid or base enter the molarity Note The acid base control is tuned with NaOH and HCI Ifyou are using acid or base with a different strength you should elaborate with the Molarity param eter to get optimal pH control Ped CUI al Function Acid Gas control Speed C 101 4 rpm Cellbag pumps Flow rate 0 0 ml min Pump unit 1 A Molarity 0 50 mol L HCL equivalent Tube 3 2 mm B inner diameter ble abd Pump unit 2 Reset total acid A Calibrate B
210. the Show details checkbox is selected This option can be used to simplify the workflow later Result The Var button changes to VAR to confirm the new variable Note If a breakpoint is defined as a variable changing the variable value in the Variable List tab when the method run is started will shift other instruction breakpoints accordingly 4 Click Change Result The variable is saved and displayed in the Text Instructions area Identifying variables in the instruction box Parameters that are defined as variables in the text method are also indicated in the Instruction Box for the selected instruction in the Text Instructions area When the instruction is shown in the Instructions field of the Instruction Box the text on the button to the left of the parameter field is displayed as VAR The button text for parameters not having associated variables is Var Instruction Box q Breakpoint Instructions lz Parameters for Heating O00 hour E Rocker Temperature setpoint 15 0 40 0 Heating ie War Heating Wen Waa L Enable heating Deviation alarm Disable heating Gas flow a Let Oot On E aa mix Allowed deviation up 0 2 5 0 E mix re pH sensor 0 5 aE pH control Allowed deviation down 0 2 5 0 pH control advanced ll a ja H DO zenzor 1 Var 1 0 C E Pi a re ii Deviation alarm delay 0 3600 control advance li p z Media control
211. to 12 Terpersture_ Chart 1g 3 to 12 Castiow Chart pet 3 to 12 wt iy yas oh Chart 1 oh 3 fo 12 90 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 2 Open and view results Note The marker will measure the curve that currently is selected if several curves are displayed in the chart The marker will have the same color as the selected curve Set marker reference You can use the vertical marker for more measurements than just the readings from a specific curve position The table below describes how to use the marker to determine Delta and Mean Y axis values Step Action 1 e Position the marker where you want to begin the measurement e Right click and choose Set vertical marker reference point Result The reference point is set to the position shown in the box in the top left corner of the chart 2 e Drag the marker to the point where you want the measurement to end Result The measured area is colored as illustrated below 2 e Read the Delta and Mean values from the box ai 2077 hour 2012 10 17 14 41 22 59 kg Delta 0 30 hour Mean 14 91 kg Min 6 51 kg Mar 22 59 kg xuri Cell Expansion System W25 User Manual 29 0646 22 AA 91 3 The UNICORN software 3 4 Evaluation in UNICORN 3 4 2 Open and view results Snapshots You can take a Snapshot of all the curve values at the marker position The table below
212. ton Stop the rocking by clicking the right hand side of the Rocking button Verification of gas flow Follow the instructions below to verify that the gas flow is functioning correctly Step Action i 2 146 Switch on the client computer Switch on the rocker and wait for the system to initialize Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Step 4 System control description 4 6 System verification Action Start UNICORN and connect to system Make sure that the desired gas supplies are connected to the system Refer to Section 5 1 5 Connect gas to system on page 166 In the Process Picture click the Compressed Air button to select compressed air or the N2 button to select N gt Enter the gas flow setpoint 0 50 in Settings Gas control Gas flow Setpoint in the Process Picture Click OK Start the gas flow by clicking the right hand side of the Gas flow button Result The gas flow starts to increase to the gas flow setpoint Fi Gas flow 0 50 L min m i Verify that the overpressure alarm is functioning by blocking the gas flow in the Gas mix tubing Result After about 5 seconds an overpressure alarm should be activated and the gas flow should stop automatically If the gas flow appears not to function correctly refer to Section 7 3 Xuri Cell Expansion W25 CBCU on page 212 Stop the gas flow by clicking the right hand side of the Gas flow button
213. trademark of ICU medical Inc Tygon is a trademark of Saint Gobain Performance Plastics Virkon is a trademark of E du Pont de Nemours and Company or its affiliates Microsoft Windows and Windows Server are registered trademarks of Microsoft Corporation Any use of UNICORN is subject to GE Healthcare Standard Software End User License Agreement for Life Sciences Software Products A copy of this Standard Software End User License Agreement is available on request UNICORN 6 2009 2014 General Electric Company 2014 General Electric Company All rights reserved First published Feb 2014 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 repre sentative for the most current information GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare UK Limited 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 Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 29 0646 22 AA 02 2014
214. tuating pH Variation in pH caused by the rocking should be less than 0 05 pH units The rocking disturbs the pH readings Check that the fiber cables are not pinched or moving excessively Place all the fiber cables in the tubing exit Stop the rocking and observe the OH readings Check that the volume in the Cellbag bioreactor is not less than the specified minimum volume Incorrect pH reading DO control Error symptom Entered pH calibration values are not correct for the bag used Check the pH label on the bag enter the correct values and perform an offset calibration The pH sensor is de graded due to long use light exposure or pres ence of substances that are harmful to the sensor like strong bases or ethanol The sensor is no longer useful for pH measure ment Automatic pH control can no longer be performed A gas bubble may be trapped on the pH sen SOT Possible cause Tap the pH sensor from the underside to remove the bubble This may require forceful manipulation and the Cellbag bioreactor may need to be partly disconnected from the tray Note The gas bubble may be very difficult to see Corrective action DO control is not start ing Some of the criteria for starting the DO control are not fulfilled Read the message in the pop up dialog If possible take action according to the information and try again Xuri Cell Expansion System W25
215. ump Description 31 CBCU Description 26 Status LED 27 Troubleshooting 212 CBCU specifications 239 Cellbag bioreactor Description 32 Illustration 33 Troubleshooting 227 Cell cultivation Tips 141 Chromatograms Insert a marker for measure ments 90 Measure using marker refer ences 91 CO Function verification 147 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Index CO mix control settings 256 Component specification CBCU specifications 239 equipment dimensions 238 equipment weight 238 Pump 25 specifications 240 Rocker specifications 239 Control cycle 119 Control mode 118 Control parameters 15 Control scheme 118 Curves Highlight in a chro matogram 90 Measure using a marker reference 91 Cycle time Acid base control 124 CO2 control 122 O2 control 129 Speed control 131 Tuning 122 129 D Dead band 131 Deviation alarm 120 DO Control mode Control scheme 118 Reading cycle 117 Start reading 117 DO control O2 control mode 129 Speed control mode 131 Transition delays 132 DO control settings general 279 O2 control mode 280 Speed control mode 281 transition delay 282 Documentation 11 DO measurement principles 115 DO mesurement settings 277 DO sensor Connect 156 287 Index Description 34 E Edit Method notes 62 Variables 68 Emergency stop Pumps 138 Equipment dimensions 238 Equipment weight 238 Evaluation module description 87 icons 87
216. urement related settings Description Calibration value high pH end of the sensor range Settings Cellbag Set from pH sensor calibra tion values View from PP RD CD Calibration value low pH end of the sensor range Settings Cellbag pH sensor calibra tion values cpHO Calibration value related to offset calibration Settings Cellbag pH sensor calibra tion values cdpH Calibration value related to resolution Settings Cellbag pH sensor calibra tion values ctemp Calibration value calibra tion temperature Settings Cellbag pH sensor calibra tion values Reading cycle time The reading cycle when pH control is off Settings pH Set pH reading cycle time Viewed when OH con trol is off Reset cali bration values at END 266 A System setting that de fines if the pH calibration values shall be reset or not when the system goes to END Note System settings pH sensor Reset cali bration values at END In the table the following abbreviations are used PP Process Picture MI Manual instruction RD Run data CD Curve data Y Yes No or Not applicable Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Output data pH measurement timer 8 Reference information 8 8 Control settings 8 8 7 pH measurement Description The time left to next pH measurement View from P
217. urn into inactive state In this state the control is not doing anything until all the necessary conditions are fulfilled again Then the control will then start again When the control inactivate a red or orange frame will be visible around the pH DO icon in the process picture A message dialog will inform why the control is inactive e An orange frame indicates that the inactivation is due to a nominal user or system action like entering sampling mode or feed harvest auto calibration e Ared frame indicates that the inactivation is due to an error Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 2 pH control 4 2 pH control In this section Xuri Cell Expansion System W25 has automated pH control In general PID parameters and cycle times do not need to be entered or tuned Moreover acid and base flow rates do not need to be calibrated or tuned Refer to Section 8 8 7 poH measurement on page 265 and Section 8 8 8 pH control on page 268 for information about pH control settings This section contains the following subsections Section See page 4 2 1 CO2 control mode 122 4 2 2 Acid Base control mode 124 4 2 3 pH control transition delays 127 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 121 4 System control description 4 2 pH control 4 2 1 CO2 control mode 4 2 1 CO2 control mode Introduction At every execution of the CO control a PID regulator computes t
218. user set weight setpoint This output data shows the current value of the ramped setpoint Auto calibration cycle The time left to start of next auto calibration hours timer Total feed The accumulated volume computed by the pump based on the estimated flow and run time Total harvest The accumulated volume computed by the pump based on the estimated flow and run time Feed calibration factor Therelation between the expected flow to RPM conver sion for selected tube diameter and the result of the cali bration Harvest calibration The relation between the expected flow to RPM conver factor sion for selected tube diameter and the result of the cali bration 1 This run data is not specifically connected to the feed or harvest waste role Instead this run data is connected to a specific pump head Look at the run data from the pump head of interest Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not applicable 286 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA Index A Acid Base control mode Dead band 126 Flow rate 125 Adjustable foot Description 18 Administration module description 41 icons 41 Alarm Deviation 120 139 Auto calibration Criteria 137 Pumps 136 B Bioreactor system Description 13 Illustration 13 Troubleshooting 207 C CAN ID CBCU Description 28 P
219. with the supplied stickers 7 Bundle the optical fiber cables and place them in the tubing exit See Illustra tions of tray and lid on page 20 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 157 5 Operation 5 1 Prepare the system 5 1 2 Prepare pH and DO sensors Step Action 8 Connect the pH sensor cable to the pH port on the CBCU front panel see below 8 Nut Cat Expormion W25 CECU pH GAS MO OUT ian gt Ps 9 Connect the DO sensor cable to the DO port on the CBCU front panel see below 2 Mure Cel Expansion W26 cecu o GAS MX QUT fin 158 xuri Cell Expansion System W25 User Manual 29 0646 22 AA 5 Operation 5 1 Prepare the system 5 1 3 Attach the Cellbag bioreactor 5 1 3 Attach the Cellbag bioreactor Instructions Follow the instructions below to attach the Cellbag bioreactor to the tray Note When using a Cellbag bioreactor that only covers half of the tray such as a 10 L bag on Tray 20 make sure to position the bag on the left side of the tray Step Action 1 Push the bag clamp opener in one upper corner of the tray downwards This opens the upper bag clamp 2 Insert the upper Cellbag rod into the opened bag clamp xuri Cell Expansion System W25 User Manual 29 0646 22 AA 159 5 Operation 5 1 Prepare the system 5 1 3 Attach the Cellbag bioreactor Step Action 3 Push the bag clamp opener upwards to secure the upper end of the Cellbag bioreactor Gently pull t
220. xpansion System W25 User Manual 29 0646 22 AA 139 4 System control description 4 4 Media control 4 44 Media control inactivation 4 4 4 Media control inactivation If any of the conditions for running the Media control is suddenly not fulfilled when running the control the control will turn into inactive state In this state the control is not doing anything until all the necessary conditions are fulfilled again Then the control will then start again When the control inactivate a red or orange frame will be visible around the Media control icon in the process picture A message dialog will inform why the control is inac tive e An orange frame indicates that the inactivation is due to sampling mode e Ared frame indicates that the inactivation is due to an error 140 Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 4 System control description 4 5 Recommended operating conditions 45 Recommended operating conditions Introduction This section includes recommendations for gas flow rocking speed and rocking angle Suitable settings for temperature pH DO CO and O are selected depending on the application and system configuration For more information about rocker control settings refer to Section 8 8 1 Rocking control on page 249 Rocking speed and angle The optimal agitation for a process is determined by the cell culture oxygen demand and the cellular shear sensitivity Rocking speed and angle should be
221. y pH control PUMP 25 2A total time ofac When the pump is running for limited time this data Y tual shot represents the total time the pump will run PUMP 25 2B actual flow The estimated flow of the pump Y Y PUMP 25 2B actual speed The pump rpm measured by the pump itself y y PUMP 25 2B accumulated The estimated total volume since last reset Y Y y volume Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 8 Reference information 8 8 Control settings 8 8 6 Pump control PUMP 25 2B time left of actu al shot Description This is counter showing how much time is left until the pump stops when the pump is running for a limited time This is the case when the pump is started manually for a limited time during calibration or when used as acid base pump by pH control View from PP RD CD PUMP 25 2B total time of ac tual shot When the pump Is running for limited time this data represents the total time the pump will run 1 Only shown in process picture when pump is used by pH or Media control 2 This curve data is not specifically connected to a pump head Instead this curve data is connected to a specific pump role acid base feed or harvest waste See curve data Total acid Total base Total feed or Total Har vest Waste Note In the table the following abbreviations are used PP Process Picture RD Run data CD Curve data Y Yes No or Not ap
222. ystem specifications of the Rocker Parameter Rocking speed control range 2 to 40 rpm 1 rocking is one complete cycle Tray 50 limited to 240 rom x angle Rocking angle control range 2 to 12 Rocking speed profile range 15 to 100 The unit relates to how much of the angular displacement function that is si nusoidally shaped 15 gives an almost constant angular velocity 100 gives a sinusoidal rocking Media weight control range Scale absolute accuracy 0 5 to 25 kg 0 050 1 of load kg Temperature sensor Pt100 Class A Temperature measurement range 2 C to 50 C Temperature control range Ambient temperature 5 C to 40 C Temperature control accuracy excl measurement error Xuri Cell Expansion W25 CBCU specifications OAS The table below he table below lists the system specifications of the CBCU Parameter Gas flow control range 0 ml min to 1000 ml min Gas flow total accuracy Process value reference flow 10 3 of read value ml min Fast fill flow Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 3 min 8 Reference information 8 2 Component specifications Parameter CO measurement accuracy 0 3 3 of read value when mixed only with air N gt CO control range 0 to 15 CO Total CO accuracy Process value ref 0 7 3 of read value CO erence CO content O measur
223. zero Tuning PID parameters and cycle time If further tuning of PID parameters and or cycle time is needed follow these steps Step Action 1 Open Tools Customize and select the tab Run Data Groups Select the Auto DO O2 P Auto DO 02 I and Auto DO O2 D and click OK Result The automatic PID parameters are shown as Run Data 2 Register the values both when approaching the new setpoint and when keeping the setpoint Tip These automatic values are a good starting point for the tuning Xuri Cell Expansion System W25 User Manual 29 0646 22 AA 129 4 System control description 4 3 DO control 4 3 1 O2 control mode Step Action 5 Open the manual instruction DO control advanced DO control advanced 02 and select Manual PID parameter mode and or Cycle time mode and enter the desired values When elaborating with a manual parameter remember that increasing the parameter will reduce the integrating effect and vice versa Instructions Manual instructions System 2 H C02 mix 02 mix pH sensor pH control H pH control advanced DO sensor DO control DO control advanced DO control advanced 02 DO control transition delays Watch parameters Other a zal Parameters for DO control advanced 02 PID parameter mode A O Auto Manual Manual P 0 00 10 00 oOo 0 20 Manual 1 100000 i 1800 Manual D 0 10000 o lt Sav

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Xuri™CellExpansionSystemW25 - GE Healthcare Life Sciences