WAVEPODTM OPERATOR MANUAL

Disclaimer of Liability GE Healthcare Bioscience BioProcess Corp. (WAVE) is not responsible for any special, indirect, punitive, incidental, or consequential damages arising out of the sale, installation, use or inability to use any product or service or any breach of warranty, under any legal theory, including lost profits, business interruption, goodwill, damage to or replacement of Equipment or property. Use restrictions are a condition of the purchase. Buyer is solely liable to ensure compliance with any regulatory requirements related to the Buyer's use of the Equipment. Any warranty granted by WAVE to the Buyer shall be deemed void if Equipment covered by such warranty is used for any purpose not permitted. Warranty, Return and Repair Policy WAVE warrants the WAVEPODTM against defects in materials and workmanship for 12 months from the date of purchase. If you discover a defect, we will, at our option, repair, replace, or refund the purchase price. Please contact the distributor from whom you purchased the Equipment for repair. NOTE: Any physical abuse of the device, or unauthorized attempts to repair or modify the device also voids the warranty. Patents US and European patents granted and pending. Wave Bioreactor, Cellbag, and Oxywell are trademarks of GE Healthcare Bioscience BioProcess Corp.

GE Healthcare Bioscience BioProcess Corp. Part # 450032 F Email : Wave.Support@ge.com Web: http://www.wavebiotech.com For USA sales and service contact: GE Healthcare Bioscience BioProcess Corp. 300 Franklin Square Drive Somerset, NJ 08873 USA Tel (732) 302 3100 Fax (732) 302 3101

WAVEPOD

Contents
Introduction 1
Manual Contents.......................................................................................................................................1 Equipment Description...........................................................................................................................1 Features.........................................................................................................................................................5 Unpacking. ...................................................................................................................................................5 Safety ..............................................................................................................................................................5

Operating Controls Navigation

Navigation ....................................................................................................................................................6 Overview Screen........................................................................................................................................7 Setup Screen. ..............................................................................................................................................8 Handling Alarms..................................................................................................................................... 11 Trends.......................................................................................................................................................... 12

Installation

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Location ...................................................................................................................................................... 13 Equipment Required ............................................................................................................................. 13 Connecting Power. ................................................................................................................................ 14 WAVEPODTM Control Modules Installation and Help Screens........................................... 15 Connecting a Base Bioreactor Unit............................................................................................... 18 Connecting Acid/Base Pumps ......................................................................................................... 19 Connecting Alarm Monitor ............................................................................................................... 20 Connecting to Data Acquistion ....................................................................................................... 20

Operating the WAVEPOD

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Base Unit Supervisory Control ......................................................................................................... 21 pH Control.................................................................................................................................................. 23 Aeration Control...................................................................................................................................... 30 Dissolved Oxygen Control.................................................................................................................. 32 Oxygen Control ....................................................................................................................................... 39 CO2 Control................................................................................................................................................ 44

Maintenance

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Routine Maintenance........................................................................................................................... 49 CO2 Sensor Adjustment ....................................................................................................................... 50 Replacement Module Parts List....................................................................................................... 52 Troubleshooting...................................................................................................................................... 53 Communications and Data Aquisition......................................................................................... 57 Connecting to PC Systems................................................................................................................. 64 Data Register ........................................................................................................................................... 65 WAVEPOD Dataset ................................................................................................................................ 68 Specifications........................................................................................................................................... 72

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Technical Reference

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Optical Dissolved Oxygen .................................................................................................................. 77

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Introduction

Manual Contents
This manual covers the installation, operation, maintenance, technical reference information and troubleshooting for the WAVEPOD instrument module manufactured by WAVE. To ensure proper operation of your WAVEPOD, please read this manual completely.

Equipment Description
The WAVEPOD instrument module combines a number of instruments for use with Wave BioreactorTM disposable cell culture devices manufactured by WAVE. It can provide dissolved oxygen measurement and control; pH measurement and control; aeration control and O2/CO2 gas blending. The WAVEPOD consists of a stainless-steel chassis with a color touchscreen that provides a simple and intuitive user interface. Data communications to Wave Bioreactor base units are provided so that the bioreactor can be integrated with the instrumentation. A built-in Ethernet port provides a digital interface for data acquisition. Seven modules are available, four of which can be installed into the WAVEPOD at a time to provide the desired functionality. A WAVEPOD is designed to be used with a single Wave Bioreactor. Each bioreactor can hold 2 CellbagsTM; one WAVEPOD is required for each Cellbag. The bioreactor can be operated from the WAVEPOD touchscreen. If space is limited, one WAVEPOD may be stacked on top of another. The built-in color touchscreen provides complete information on installation, configurations, connections, utilities, communications, pumps and alarms.

WAVEPOD

BAY 1 pH BAY 3 AIRPUMP BAY 2 DO BAY 4 O2, CO2 or O2+CO2 Touchpanel

Ethernet Status

Figure 1. WAVEPOD Instrument Module.

BAY 1 pH

BAY 3 AIRPUMP

BAY 2 DO

BAY 4 O2, CO2 or O2+CO2

Figure 2. WAVEPOD Module Layout.

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Figure 3. Rear Panel Connectors and Controls.

pH CONTROLLER
The pH control module enables on-line control of pH in the Cellbag. This module is installed in BAY 1. A setpoint controller is provided to power BASE and ACID pumps on and off to maintain pH. The controller can operate WAVE PUMP20 units or user-supplied pumps. Alternatively, the CO2 concentration can be controlled (requires CO2 CONTROL option) to maintain pH.

DO (DISSOLVED OXYGEN) CONTROLLER

The DO control module enables on-line control of dissolved oxygen (DO) in the Cellbag. The DO control module is located in BAY 2. The controller monitors dissolved oxygen using an optical sensor and can be configured to vary the rocker speed to maintain the desired dissolved oxygen concentration in the Cellbag. Alternatively the controller can control the % O2 in the aeration gas to the Cellbag (requires O2 CONTROL option).

AIRFLOW CONTROLLER
The airflow control module is located in BAY3. It can be used as a standalone module for Cellbag aeration or as a conditioned gas supply to a WAVE Bioreactor20/50EHT unit. Air is aspirated from the AIR IN port on the module panel and pumped out the AIROUT port. The AIR IN port can be left unconnected to use ambient air or process air (0.2 BAR max) can be connected. The airflow is measured by a thermal mass flow sensor and regulated by a flow control valve operated by an internal controller. The AIRFLOW control module is required if CO2 or O2 modules are to be used.

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CO2 CONTROLLER
CO2 concentration is measured and controlled by the CO2 control module located in BAY4. The gas composition is measured by a Non-dispersive infrared (NDIR) CO2 sensor using a flow-through cell. The internal controller regulates CO2 gas to mix with air to produce the desired CO2 concentration in the gas to the Cellbag headspace. The CO2 sensor is temperature compensated (0 50 C). Pressure changes can influence the reading however, the gas flow is carefully regulated to maintain constant pressure in the CO2 sensor cell regardless of AIROUT delivery pressure or minor fluctuations in CO2 inlet pressure. Since the CO2 controller uses a CO2 measurement to control the composition, it is not necessary to use pure CO2 gas for control.

O2 CONTROLLER
O2 concentration is measured and controlled by the O2 control module located in BAY 4. The gas composition is measured by a unique zirconium oxide sensor. The oxygen specific chemistry produces a signal proportional to the oxygen present in the flow through cell and microprocessor circuitry provides a linear and stable signal. The O2 sensor can be adjusted using ambient air and/or span gases. Unlike electrochemical sensors, O2 sensor does not degrade when not in use. Under typical operating conditions, the sensor should last 10 years. O2 using a flow-through cell. The internal controller regulates how much O2 gas to mix with air to produce the desired O2 concentration to the Cellbag headspace. The O2 sensor is temperature compensated (0 50 C). Pressure changes can influence the reading; however, the gas flow is carefully regulated to maintain constant pressure in the O2 sensor cell regardless of AIROUT delivery pressure or minor fluctuations in O2 inlet pressure. Since the O2 controller uses an O2 measurement to control the composition, it is not necessary to use pure O2 gas for control.

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Features
Compact design provides all major bioreactor instrumentation Large color touchscreen provides easy user access Integrates bioreactor to provide a unified control unit with advanced control capabilities Easy expansion using self-configuring plug-in control modules Ethernet output for data acquisition Date-stamped alarms stored in non-volatile memory for easy troubleshooting Integrated control for acid/base pumps Alarm contacts for chart recorders and remote monitoring systems Available in right or left facing versions Stackable (up to 2 high)

Unpacking
Verify that all parts are present. You should have:
WAVEPOD with all ordered modules installed US Power cord EU Power Cord DOOPT Orange Extension Cable (when DO option is installed). Communication Cable Air-Line Assembly, POD to Cellbag Air-Line assembly, POD to bioreactor base Calibration Record Operator Manual P/N 450032 POD CD

Other accessories may be present. Please check the packing slip against your order. Contact WAVE immediately if any parts are missing or damaged.

Safety
The WAVEPOD should not be used for any application other than specified by the manufacturer. If the WAVEPOD is used in a manner not specified by WAVE, the protection provided by the equipment may be impaired and the warranty may be voided. Equipment should not be used in hazardous atmospheres or with hazardous materials for which the equipment was not designed. This equipment is designed for indoor use only.

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Operating Controls Navigation

Navigation
The WAVEPOD is controlled and navigated though the touchscreen controller. There are several types of screens used in the control and operation of the WAVEPOD. Overview screens (light blue) contain the basic functions to operate a bioreactors pH, Dissolved Oxygen (DO), Oxygen (O2), Carbon Dioxide (CO2), and Aeration. Control screens (yellow) contain detailed setup controls for each function and are only accessible by a password. An ALARM LOG screen is accessed through the SYSTEM INFO screen that provides a record of all alarms and permits acknowledgement of alarms. Figure 4 illustrates the relationship between screens. The BACK button permits returning to a previous screen.

Figure 4. WAVEPOD Control Screens.

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Overview Screen
After initialization, the unit will display the OVERVIEW screen: The actual OVERVIEW screen you see may be slightly different depending on the options installed.

Figure 5. Overview Screen. The left side of the screen shows data (if connected) from the BASE unit connected to the WAVEPOD. The right side of the screen displays the instruments in the WAVEPOD itself. Red numerals in black boxes are measured process variables (PV). Grey boxes are function buttons. Pressing one of these will bring up the corresponding detail screen. Setpoints are displayed as black numerals in grey boxes. Pressing on a setpoint (SP) box will bring up a numeric keypad. If any alarm conditions exist, the ALARM button is displayed flashing at the bottom of the screen. If the alarm is new and has not been acknowledged, then this ALARM button will be flashing. Pressing the ALARM button will bring up the ALARM LOG screen. See the Handling Alarms section for instructions on what to do with alarms. Pressing the MENU button brings up the SYSTEM INFO screen which allows you to access equipment setup and other functions. Pressing the HELP button brings up a series of help screens that are useful for setup and troubleshooting. Instrument modules plugged into the WAVEPOD are detected automatically and will appear on the display and setup screens. Various options can be set by the user. The options are protected from unauthorized access by a password. Password 2050.

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Setup Screen
To access the options press the MENU button from the OVERVIEW screen. This will bring up the following SYSTEM INFO screen:

Figure 6. System Information Screen. The SYSTEM INFO screen provides:

WAVEPOD serial number PLC version number TP (touchpanel) program version Compile date Pod Bay Status Alarms Bay/Base Alarm Acknowledgement Communication Status Base Status

It also provides an overview of current alarm status for the POD and any connected base unit. Alarms may be reviewed by pressing REVIEW ALARMS. Pressing SETUP will bring up a password entry screen. This password prevents unauthorized personnel from accessing SETUP parameters. The default password is 2050.

Figure 7. Setup Access Password Screen. After the correct password is entered a PROCEED button will be displayed. Press this button to access the SETUP screen:

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NOTE: You may see a different SETUP screen depending on the specific options installed in your WAVEPOD

Figure 8. Setup Screen.

OPTIONS
Pressing the OPTIONS button will bring up the OPTIONS screen:

Figure 9. Options Screen. Select type of BASE unit: BASE type: Press on this window with the model showing repeatedly until the appropriate BASE unit model is displayed. Available options are: 2050EHT_SINGLE 2050EHT_DUAL 2050EHT_CO2 2050EHT_O2 NONE Select NONE if no base unit is connected. Default: NONE. Monitoring options can be set: RESET: Press RESET to accept selected BASE unit.

WAVEPOD

Auto Start: Setting these to ON will cause the appropriate device (POD or BASE) to resume operating on power up. All controls operate exactly as before power was lost or shut down. This is useful if it is desired that the unit resume operation after power failure. Default: ON. Auto Ack: (Automatic acknowledge): Setting these to ON causes any alarms from the appropriate device to be acknowledged automatically. The alarms are still recorded. Default: OFF. Alarm Beeper: Setting this to ON causes any alarms to trigger the internal audible beeper. Turning the beeper OFF prevents the alarms from being acknowledged. Default: ON.

SETTING CLOCK
Pressing SETCLK will bring up the SET REAL TIME CLOCK screen:

Figure 10. Real Time Clock Screen. The real-time clock can be set by pressing on the time parameters. A 24 hour (military time) clock is used.

CLEAR ALARM LOG

Pressing the CLEAR ALARM LOG button clears the stored alarm log. The alarm log cannot be retrieved if accidentally deleted using this function. CAUTION When the CLEAR ALARM LOG button is pressed, all alarms are cleared and cannot be restored.

Figure 11. Setup Screen.

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CONTROLLER PARAMETERS
Other buttons on the SETUP screen are used to access various controller parameters depending on the installed options. These are described in the respective controller Operating sections.

Handling Alarms
All alarms are time-stamped to facilitate troubleshooting. The troubleshooting section in Maintenance includes a list of all alarms and troubleshooting hints. ALARM BEEPER The beeper sounds every time a new alarm occurs. The beeper can be disabled (ALM BEEPER OFF) from the SETUPOPTIONS screen. When a new alarm occurs, a red flashing ALARMS button appears on the bottom of the displayed screen. This indicates that the alarm has not been acknowledged. An audible beeper sounds (if Alarm Beeper is enabled) alerting the operator that a new alarm has occurred. Press the ALARMS button to access the alarm history screen. This action also acknowledges the alarm and makes the ALARMS button turn yellow and stop flashing. The audible alarm will also be silenced. The ALARMS button remains visible until all alarms have cleared. On the ALARM LOG screen each alarm is recorded along with the time it occurred or cleared. Each alarm entry has the alarm number, description and status. The alarm history is retained on power failure or shutdown. The last 1000 alarms are retained. Use the up and down scroll arrows to page through the alarm log. The alarm history can be cleared from the SETUP menu (MENU-> SETUP-> CLEAR ALARM LOG. Once cleared, the alarm log cannot be recovered. To review an alarm, touch the alarm message. This brings up a pop-up window with more information about the alarm. The alarm message turns blue indicating that alarm has been viewed.

Figure 12. Alarm Log Screen.

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RED MESSAGE: GREEN MESSAGE BLUE MESSAGE

Alarm not acknowledged Alarm condition cleared Alarm has been reviewed

If any alarm condition exists, then the UNIT ALARM contact relay is set. Refer to Maintenance section for wiring details. The UNIT ALARM is cleared once all alarm conditions have cleared. Many alarm conditions and limits can be set by the user. Use the SETUP screen to set parameters. The ALARM LOG can be reviewed at any time by pressing the ALARMS button from the OVERVIEW screen or by pressing the REVIEW ALARMS button from the SYSTEM INFO screen.

Figure 13. System Information Screen.

Trends
Most of the CONTROL screens have a TREND button. Pressing the associated TREND button brings up a specific trend for the associated variable. Trends contain data for the last 10 minutes of operation and are useful for troubleshooting. The WAVEPOD does not maintain long-term trends. Figure 14. Typical Trend Screen.

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Installation

Location
The WAVEPOD should be physically installed next to the Wave Bioreactor or as close as possible. Two configurations are available for the WAVEPOD, left and right side module locations. Place the WAVEPOD on the proper side of the bioreactor to allow easy installation of tubing and cables. DO NOT place the WAVEPOD next to a heating register, air duct or in direct sunlight as control operations may be affected. One WAVEPOD may be stacked one on top of the other. NOTE: One WAVEPOD is required for each Cellbag installed on a Wave Bioreactor.

Equipment Required
Depending on the options installed, the items listed below are recommended to facilitate the installation of the WAVEPOD. Some of the items are facility sources and other items can be purchased from WAVE. Facilities Oxygen (O2) supply source pressure regulated to 0.7 to 1 bar (10-15 psig) Carbon Dioxide (CO2) supply source- pressure regulated to 0.7 to 1 bar (10-15 psig) Acid pump- WAVE Model PUMP20 (or customer supplied pump) Base pump- WAVE Model PUMP20 (or customer supplied pump) pH Probe DOOPT Probe CIDEX disinfectant solution for pH probes

Equipment and Materials

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Connecting Power
1. Connect the instrument to AC power (90 to 240 VAC, 50/60 Hz) using the provided power cord. The power rating is 3 Amperes. The power inlet is located on the rear of the instrument. NOTE: Protection provided by the equipment may be impaired if used in a manner not specified by the manufacturer. 2. Turn on power to the unit. The Ethernet indicator should light up and the following startup screen should be displayed within 20 seconds: BASE not found message is displayed initially until a Base unit is connected to the WAVEPOD. This is normal. 3. After a few seconds the OVERVIEW screen will be displayed:

Figure 15. Initial Displays-Normal and Without Base Connected. NOTE: You may see a different OVERVIEW screen depending on the specific options installed in your WAVEPOD.

Figure 16. Overview Screen Display.

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WAVEPOD Control Modules Installation and Help Screens

With the Overview screen displayed, press the HELP button. The WAVEPOD HELP screen is now visible. This screen permits access to five additional screens as shown.

Figure 17. WAVEPOD Help Screen. Press the INSTALLATION button. A new screen appears illustrating where each module is assigned a specific bay in the WAVEPOD as shown below.

Figure 18. Installing Instruments Screen.

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If you purchased modules with your WAVEPOD, the modules will be pre installed in the unit. If you have empty bays and wish to install the corresponding instrument proceed as follows:

1. Power off the WAVEPOD. 2. To install a new module, remove the 4 fastening screws from the appropriate bay cover plate on the side of the unit. See Figure 18. 3. Insert the new module. Make sure that it is fully seated. When installing a CO2 or O2 module make sure that the gas flow tubes are properly connected to the AIRPUMP module located in bay 3. All tubing ends are color coded. 4. Secure the module firmly with the 4 fastening screws. 5. Power up the instrument. The new module is configured automatically and shows up on the INSTALLING INSTRUMENTS screen when selected. 6. Press the BACK button on the INSTALLING INSTRUMENTS screen, then, press the CONNECTIONS button. A HARDWARE HELP screen appears. This screen is a schematic diagram illustrating the hardware connections as shown in Figure 19. Use this as a guide.

Figure 19. Hardware Help Screen. To view the other screens, press the BACK button on the current screen, or select the appropriate screen from the WAVEPOD HELP screen.

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Figure 20. Connecting Utilities Screen.

Figure 21. Acid Base Pumps Screen.

Figure 22. WAVEPOD Communications Screen.

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Connecting a BASE Bioreactor Unit

IMPORTANT The WAVEPOD can only connect to a single base unit. The baud rate on the base unit is set to 9600. DO NOT CONNECT anything to the second dataport on the BASE unit as communication protocol will be violated. A Wave Bioreactor BASE unit can be connected to the WAVEPOD to provide an integrated control platform (WaveCLUSTER). Additional units may be connected as shown in section on Ethernet Connections. The WAVEPOD is mainly designed to interface with the SYSTEM20/50EHT bioreactor. To connect a BASE unit: 1. Locate the supplied WAVEPOD-BASE interconnect cable. Connect one end to the port labeled BASE20/50EHT on the rear panel of the WAVEPOD. 2. Connect the other end of the 6 pin RJ12 cable to DATAPORT 1 jack on the BASE unit. See Figure 23. Do not connect to DATAPORT 2.

DO NOT CONNECT TO DATA PORT 2

WAVEPOD Rear Panel

SYSTEM20/50EHT Base Unit Rear Panel

Figure 23. WAVEPOD Connection to a SYSTEM20/50 EHT Unit.

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3. Navigate to the SETUP MENU. Select the OPTIONS screen and select the type of BASE unit. 4. Now cycle power OFF and ON to initiate communications with the connected BASE unit. After 20 seconds the SYSTEM INFO screen appears. Then, if the BASE unit is connected properly, the OVERVIEW screen with BASE data appears on the left side of the screen as shown in Figure 24. If the BASE unit fails to communicate with the WAVEPOD, a BASE COMM FAILED message appears as shown in Figure 25.

LOST COMMUNICATIONS If the BASE unit is powered off or the cable to the WAVEPOD is disconnected, then communications may not resume on power up or reconnection. The OVERVIEW screen shows a RETRY button. Pressing the RETRY button resets the WAVEPOD and restarts communications.

Figure 24. Overview Screen Shown with Base2050EHT Selected.

Figure 25. Overview Screen Shown with Base Communication Failure.

Connecting Acid/Base Pumps

The WAVEPOD can operate acid and base pumps for pH control. Two DB9 jacks are located on the rear panel. These jacks are designed to interface with WAVE PUMP20 modules or pumps you choose. Using the cables supplied with the pump modules, connect the acid pump to the jack marked PUMP1 and the base pump to the jack marked PUMP2. If using pumps other than the PUMP20 units, you need to make a suitable cable based on the jack pinout. The pinout is provided in the Communication and Data Acquisition section. PUMP1 PUMP2 acid pump base pump

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Connecting Alarm Monitor

A WAVEPOD may be connected to an external monitoring system to alert in the case of an alarm condition. A dry contact is provided on the rear panel. This contact will change state NO and NC if any alarm occurs or if power to the WAVEPOD is lost. Jack pinout is provided in the Communication and Data Acquisition section.

Connecting to Data Acquisition

A RJ45 jack is provided to connect the WAVEPOD via Ethernet to external data acquisition and supervisory control. A standard Ethernet cable (not provided) is used to connect this jack to a standard Ethernet hub or switch. The pinout is provided in the Communication and Data Acquisition section.

Figure 26. WAVEPOD Rear Panel Connections.

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Operating the WAVEPOD

Base Unit Supervisory Control

A Wave Bioreactor BASE unit such as a SYSTEM20/50EHT can be remotely controlled from the WAVEPOD. If the BASE is connected and operating, overview data are displayed on the left side of the OVERVIEW SCREEN:

NOTE: The BASE screen may appear different depending on the type of base connected to the WAVEPOD.

Press the 2050EHT (or other BASE ) button to bring up the control screen for the base:

NOTE: On BASE units equipped with optional weight perfusion control, the BASE CONTROL screen displays WEIGHT kg and TREND only. Control functions are made at the BASE unit.

Figure 27. Base 20/50EHT Control Screen.

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Rocking Speed: The actual rocking speed is shown in the box labeled SPEED rpm. Speed Setpoint: The rocking setpoint is shown in the box labeled SP. Pressing the setpoint brings up a data entry keypad. You can enter a new setpoint and press the ENT key to accept the new setpoint. The keypad does not permit data entry outside a preset range. Range 2 to 40 rpm. Rock angle: The rock angle is displayed. It can be changed by pressing and entering the new angle using the pop-up keypad. Range 2 to 12 degrees. Rocker ON/OFF: Pressing this button starts the rocking. The button changes to green and displays ON. Pressing it again stops the rocker and the button changes to red and displays OFF. The rocker will move to the preset SAMPLE angle. Airflow PV (Process Variable): The actual measured airflow is shown in the box labeled lpm. Airflow Setpoint: The airflow setpoint is shown in the box labeled SP. Pressing the setpoint brings up a data entry keypad. You can enter a new setpoint and press the ENT key to accept the new setpoint. The keypad does not permit data entry outside a preset range. Range 0 to 1.0 lpm. Airflow ON/OFF: Pressing this button starts the airflow controller. The button changes to green and displays ON. Pressing it again stops the airflow control and the button changes to red and displays OFF. Weight Kg: When an optional perfusion control is installed in a BASE unit, the screen will display WEIGHT kg and a TREND button. Perfusion control cannot be made from the WAVEPOD. NOTE: Most commonly used BASE functions can be accessed from the WAVEPOD. Trends: The rocking speed and other parameters are graphed over a 10 minute period. Press the appropriate TREND button to view the graph. BACK: Pressing the BACK button returns to the OVERVIEW screen.

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pH Control
FUNCTION
The pH control module enables on-line control of pH in the CellbagTM. A setpoint controller is provided to power BASE and ACID pumps on and off to maintain pH. The controller can operate WAVE PUMP20 units or usersupplied pumps. Alternatively, the CO2 concentration can be controlled (requires CO2 CONTROL option) to maintain pH. The appropriate module must be installed in BAY1. Regardless of the sensor used, the pH control action is identical.

NOTE: The pH can also be adjusted using buffers. Remove the probe from the Cellbag. Place the probe in sterile buffer pH 7 and adjust the SPAN knob on the pH module. Place the probe in buffer pH 4 and adjust the SLOPE on the pH module. This operation must be done aseptically, and then the probe can be reinserted into the bag.

ELECTRODE pH SENSOR
The pH module requires special combination electrode pH probe made by WAVE (PHPROBE) that can be inserted into a luer fitting provided on the Cellbag. These pH probes are provided sterile and must be inserted in an aseptic manner. They are also available sterilized and preinstalled in the Cellbag. The pH probe may be reused, resterilized chemically by complete immersion for a minimum of ten (10) hours in CIDEX (activated dialdehyde solution glutaraldehyde 2.4%). Reinstall probe aseptically.
CIDEX is a registered trademark of Advanced Sterilization Products, a Johnson & Johnson Company.

MEASUREMENT THEORY
pH is measured using an ultra-miniature combination glass electrode. This electrode is capable of long-term stable operation necessary for cell culture applications. NOTE: pHPROBE is packaged in a 3M NaCI 40% glycerol storage solution. Probes may be used directly from storage or the storage solution may be removed via a sterile wipe or rinsing with a sterile buffer of choice. DO NOT TOUCH THE PROBE TIP.

INSTALLATION AND ADJUSTMENT

1. Power up the instrument. Connect the pH probe using the supplied cable to the jack on the front panel of the pH module. Since the WAVE pH probes are very consistent in performance from probe to probe, it is usually not necessary to adjust using two buffers. Instead, a single point adjustment using media and a single grab sample pH measurement is usually sufficient. 2. Fill the Cellbag with media and allow to equilibrate. Measure the pH by sampling the Cellbag and using an external calibrated pH meter. Adjust the SPAN potentiometer until the reading on the controller matches the grab sample reading. The SLOPE is adjusted at the factory and should not need to be readjusted. 3. The pH probe is now ready for use.

pH probe

pH module

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pH Control Screen
Pressing the pH button (OVERVIEW screen) brings up the pH CONTROL screen.

Figure 28. pH Control Screen. pH PV (Process Variable): The measured pH is shown in the box labeled pH. pH Setpoint: The pH setpoint is shown in the box labeled SP. Pressing the setpoint brings up a data entry keypad. You can enter a new setpoint and press the ENT key to accept the new setpoint. The keypad does not permit data entry outside a preset range. Range is 5 to 9 pH units. pH ON/OFF: Pressing this button starts pH control. The button changes to green and displays ON. Pressing it again disables pH control and the button changes to red and displays OFF. CONTROL MODE: ACID/BASE or CO2/BASE - the pH controller can operate in one of two modes 1. Control by ACID/BASE pumps; or 2. Control by CO2 concentration. In the latter mode, the CO2 setpoint is remotely controlled, so that if the pH rises above setpoint the CO2 concentration of the gas to the Cellbag headspace is increased. If the pH drops below setpoint, then the CO2 level is decreased. If the CO2 reaches a minimum user-set limit, then the controller can automatically trigger a BASE pump to maintain pH. Trends: The pH is graphed over a 10 minute period. Press the TREND button to view the graph.

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pH Control Setup
pH controller parameters can be set from the SETUP menu -> pH. Access is restricted by password. Password 2050. pH parameter screen:

Figure 29. pH Control Setup Screen. Setting Deviation Alarms: Dev Alarm: If this parameter is set to OFF, then deviations from the setpoint will not be monitored. Default: On. Dev Limit: Set the pH deviation alarm deadband. A deviation alarm is triggered if the pH is different from the pH setpoint by +/- this amount. The deviation alarm is only active if DEV alarm is ON and the pH controller is ON. Range is 0.0 to 1.0 pH unit, Default: 0.05 pH. pH controller options: DBAND: Control action takes place if the pH is within the deadband. The deadband is defined +/- the setpoint. Increasing the setpoint will result in coarser control. Making the deadband too narrow will result in excessive chatter and poor control. Range is 0.0 to 1.0 pH unit, Default: 0.1 pH. CTIME: After each control action, the controller must wait for a defined cycle time (CTIME) in order to let the acid/base or CO2 change to result in a stable pH reading. If the CTIME is set too short, then the controller will overshoot. If it is set too long, then the pH may not be controlled properly and drift. The CTIME can be set by the user to match process requirements and acid/base strength. Range is 10 to 300 seconds, Default: 120 seconds.

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Available Control Schemes:

Two control schemes are defined, either can be selected by the user: ACID/BASE CONTROL: Change in pH outside the deadband causes the acid or base pump to come on as needed. The amount of time the pumps are on depends on the magnitude of the pH difference and the GAIN. The pump then stays off for the remainder of the cycle time (CTIME parameter). The gains can be adjusted to compensate for the buffering capacity of the media and the acid/alkali strength. ACID GAIN: Higher gain results in the ACID delivery pump staying on a larger fraction of the cycle time for a given pH error (pH pH setpoint). Range is 1-999, Default: 100. BASE GAIN: Higher gain results in the BASE delivery pump staying on a larger fraction of the cycle time for a given pH error (pH pH setpoint). Range is 1-999, Default: 100. CO2/BASE: The controller responds to change in pH outside the deadband by changing the CO2% setpoint. This option requires that the CO2 control module by installed, supplied with CO2 gas and be turned on. The AIRPUMP must also be ON. A low pH will result in a reduction in CO2 concentration. The concentration is adjusted by CO2STEP every cycle time to give the headspace sufficient time to equilibrate. The maximum and minimum CO2% concentration in the gas supplied to the headspace can be set by the user. An alarm is set when either of these limits are reached. If the CO2% is reduced to the minimum concentration then the BASE pump control is activated to continue pH control as needed. The amount of time the pump comes on depends on the magnitude of the pH difference and the GAIN. The pump then stays off for the remainder of the cycle time (CTIME parameter). The gains can be adjusted to compensate for the buffering capacity of the media and the acid/alkali strength. CO2STEP: This is the amount by which the controller changes the CO2 setpoint at each cycle. Range is 0.0 to 2.0% CO2, Default: 0.5%CO2. MINCO2: The controller does not reduce the CO2 setpoint below this limit. Range 0.0 to 15.0% CO2, Default: 0 MAXCO2: The controller does not increase the CO2 setpoint above this limit. Range 0.0 to 15.0% CO2, Default: 15%.

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OPERATION OF pH CONTROLLER
The pH controller provides on-line pH monitoring and control of the Cellbag disposable bioreactor. The controller will turn acid and base pumps on and off as needed. Alternatively the pH controller can change the CO2 concentration in the gas to the Cellbag.

Control
BASE

ACID

acid pump

base pump Wave Bioreactor

pH probe

CHANGING THE pH SETPOINT

The pH setpoint determines the pH that should be maintained in the bioreactor. To change the setpoint: 1. Press the pH SP display box on the pH control screen. 2. Enter the desired setpoint using the popup keypad. Press ENT to accept the new value; CLR to clear the display and ESC to abort data entry.

SWITCHING pH CONTROL ON/OFF

The pH controller can be turned on or off by pressing the CONTROL button on the pH control screen. GREEN RED control ON - control OFF

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ACID/BASE OPERATION
For stable pH control it is necessary to cycle the addition pumps so that the acid or base has sufficient time to mix properly. The pH controller has a preset factory cycle time of 10 seconds. Part of this cycle is the ON time when the acid or base pump is energized. For the remainder of the cycle, the pumps are OFF and the controller waits until the cycle time is complete. If, at the end of the cycle, the pH is still not at the setpoint, the controller initiates another ADD/WAIT cycle. If the pH is at the setpoint, no further action is required. A small dead band around the setpoint is used to prevent excessive pump chatter. The cycle time can be adjusted if the system requires more mixing time. The amount of acid or base added per cycle is proportional to the difference between the setpoint and the actual pH. The GAIN constant adjusts the sensitivity. This GAIN can be increased if insufficient acid or base is being added per cycle. It can also be decreased if very concentrated acid or base is being used and significant overshoot is observed.

CONNECTING THE BASE PUMP

The BASE PUMP is used to add base to raise the pH in the Cellbag. It can be switched on and off automatically by the pH controller to maintain a constant pH equal to the setpoint. 1. Switch power on to the controller. Select the ACID/BASE control mode (SETUP menu). Set pH controller parameters as desired. 2. A peristaltic base pump such as the WAVE PUMP20 can be controlled by the pH controller. Plug the PUMP20 base pump to the jack marked PUMP2 on the rear panel of the WAVEPOD using the supplied cables. 3. Make sure that the pump direction is correct and turn all local switches on. Route the base tubing through the pump and connect to the Cellbag container. Open all valves. 4. Set the desired pH setpoint on the controller and press the pH controller ON button to activate acid/base control. 5. The BASE pump will turn on whenever the pH drops deadband units below the pH setpoint.

CONNECTING THE ACID PUMP

The ACID PUMP is used to add acid to lower the pH in the Cellbag. It can be switched on and off automatically by the pH controller to maintain a constant pH equal to the setpoint. 1. Switch power on to the controller. Select the ACID/BASE control mode (SETUP menu). Set pH controller parameters as desired. 2. A peristaltic acid pump such as the WAVE PUMP20 can be controlled by the pH controller. Plug the PUMP20 acid pump to the jack marked PUMP1 on the rear panel of the WAVEPOD using the supplied cables. 3. Make sure that the pump direction is correct and turn all local switches on. Route the acid tubing through the pump and connect to the Cellbag container. Open all valves.

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4. Set the desired pH setpoint on the controller and press the pH controller ON button to activate acid/base control. 5. The ACID pump will turn on whenever the pH rises deadband units above the pH setpoint.

CO2/BASE OPERATION
Alternatively, it is possible to control the pH of suitably buffered media by changing the CO2 concentration of the headspace gas in equilibrium with the media. This second control mode (selectable form the SETUP menu) will make step changes in CO2 concentration to maintain pH. If the CO2 setpoint is reduced to a user-defined minimum limit, then the controller will automatically shift to BASE pump control as described earlier.

CONNECTING THE BASE PUMP

The BASE PUMP is used to add base to raise the pH in the Cellbag in the event it cannot be adjusted anymore by lowering CO2 concentration in the headspace. It can be switched on and off automatically by the pH controller to maintain a constant pH equal to the setpoint. 1. Switch power on to the controller. Select the CO2/CO2MIX control mode (SETUP menu). Set pH controller parameters as desired. 2. A peristaltic base pump such as the WAVE PUMP20 can be controlled by the pH controller. Plug the PUMP20 base pump to the jack marked PUMP2 on the rear panel of the WAVEPOD using the supplied cables. 3. Make sure that the pump direction is correct and turn all local switches on. Route the base tubing through the pump and connect to the Cellbag container. Open all valves. 4. Set the desired pH setpoint on the controller and press the pH controller ON button to activate CO2/base control. 5. The BASE pump will turn on whenever the pH drops deadband units below the pH setpoint and the CO2 controller setpoint is at preset minimum limit.

CONNECTING THE CO2 CONTROLLER

The CO2 controller can be given a remote setpoint to maintain constant pH in the Cellbag. 1. Switch power on to the controller. Select the CO2/BASE control mode (SETUP menu). Set pH controller parameters as desired. 2. Connect CO2 gas to the CO2 controller (must be installed in WAVEPOD). Toggle CO2 control to ON to turn on CO2 control. Toggle AIRPUMP control to on to turn on the airpump 3. Set the desired pH setpoint on the controller and press the pH controller ON button to activate remote setpoint CO2 control. 4. The CO2 setpoint will be adjusted up or down each cycle time if the pH is outside the control deadband.

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Aeration Control
Pressing the O2 MIX or CO2MIX button (OVERVIEW screen) brings up the O2MIX or CO2MIX CONTROL screens. The airflow control is located on the right side of either of these control screens:

Figure 30. O2 Mix Control Screen. NOTE: In LOCAL mode the outlet pressure of the AIROUT from the WAVEPOD is monitored and generates an alarm if it exceeds 5 inches of water. The pressure monitor is disabled in REMOTE mode as the BASE unit monitors gas pressure to the Cellbag. Figure 31. CO2 Mix Control Screen.

LOCAL OPERATION
If a BASE unit is not connected to the WAVEPOD or the selected BASE unit does not have airflow control, then the WAVEPOD automatically operates in a LOCAL mode: Airflow: The actual airflow in liters/minute is shown in the box labeled lpm. Airflow Setpoint: The airflow setpoint is shown in the box labeled SP, below the MIX OUT display. Pressing the set point brings up a data entry keypad. You can enter a new setpoint and press the ENT key to accept the new setpoint. The keypad does not permit data entry outside a preset range. Range is 0 to 1.0 lpm. AIRPUMP ON/OFF: Pressing this button starts the airpump and airflow control. The button changes to green and displays ON. Pressing it again
30

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disables the airpump and airflow control. The button changes to red and displays OFF. Trends: The airflow is graphed over a 10 minute period. Press the TREND button to view the graph. REMOTE OPERATION If a BASE unit with internal airflow control is connected to the WAVEPOD, then it can control the air (or mixed gas drawn from the WAVEPOD). Whenever the BASE AIRpump is switched ON, this mode becomes active and the legend REMOTE is displayed:

Figure 32. CO2 Mix Remote Base Control Screen. In REMOTE operation, the displayed airflow and airflow setpoint are the settings from the connected BASE unit. The airflow generated from the WAVEPOD is automatically set to a preset value greater than used by the BASE. If the BASE is disconnected, then the REMOTE AIRFLOW indicator displays a blank as the remote connection has been severed. Check the cables and reconnect. Cycle power to the WAVEPOD to revert the unit back to LOCAL AIRFLOW mode. BACK: Pressing the BACK button will return to the OVERVIEW screen.

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Dissolved Oxygen Control

DISSOLVED OXYGEN CONTROL SCREEN
DO Probe bar graph indicates the condition of the DO probe. When the bar graph is green, the probe is functional. When the green bar reaches the bottom, the graph turns red indicating the probe should be replaced Pressing the DO button (OVERVIEW screen) brings up the DO OPTICAL control screen:

Figure 33. DO Optical Screen. DO PV (Dissolved Oxygen Process Variable): The actual DO concentration in % saturation is shown in the box labeled %sat. DO Setpoint: The DO setpoint is shown in the box labeled SP. Pressing the setpoint brings up a data entry keypad. You can enter a new setpoint and press the ENT key to accept the new set point. The keypad does not permit data entry outside a preset range. Range is 0 to 100 +/- 0.5 %sat. DO ON/OFF: Pressing this button starts DO control. The button changes to green and displays ON. Pressing it again disables DO control and the button changes to red and displays OFF. CONTROL MODE: Rocking speed or O2% The DO controller can operate in one of two modes 1. RPM where the controller changes the BASE unit rocking rate to keep the DO at setpoint; or 2. Control by O2 concentration where the O2 setpoint is remotely controlled so that if the DO is below setpoint, then the O2 concentration in the gas to the headspace is increased. Conversely, a high DO concentration causes the controller to reduce the O2 concentration. T/COMP (Temperature Compensation): Pressing this button brings up a keypad where you can manually enter the temperature of the DO probe. This is used for temperature compensation. SCAN: Pressing this button brings up a keypad where you can set the time between successive DO readings. A longer scan time increases probe life but very slow scan time may not be fast enough for control. Range 1-99 seconds, Default: 10 seconds. DETAIL: Pressing DETAIL brings up a pop-up window displaying various probe parameters, such amplitude and phase of the signal. This display is useful for troubleshooting.

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CALIB: Pressing CALIB brings up a probe adjustment screen. Trends: The DO is graphed over a 10 minute period. Press the TREND button to view the graph.

FUNCTION
The DO control module enables on-line control of dissolved oxygen (DO) in the Cellbag. The controller monitors dissolved oxygen using an optical sensor and can be configured to vary the rocker speed to maintain the desired dissolved oxygen concentration in the Cellbag. Alternatively the controller can control the % O2 in the aeration gas to the Cellbag (requires O2 CONTROL option).

USING THE DOOPT PROBE

Remove a DOOPT-PROBE (dissolved oxygen optical probe) from its packaging. You can leave the probe in its protective tube until an adjustment is required. Connect the probe to the jack on the side panel. Make sure that the ridge on the plug is lined up with the notch on the jack. Push the plug in and twist to lock it in position. From the overview screen, press the DO OPT button. The DO display should show a DO reading. Press DETAIL to verify the signal quality. The error code should be 0, if not, refer to the Troubleshooting section of this manual for additional error code descriptions. CAUTION: DO NOT TOUCH THE TIP OF THE PROBE WITH YOUR FINGER OR ANY OBJECT. DO NOT ATTEMPT TO CLEAN OR DRY THE TIP WITH A CLOTH. SIMPLY SHAKE DRY.

ADJUSTMENT BEFORE USE

Remove the protective tube from the probe. Adjustment requires a two-point method: 1. Zero oxygen Open a new packet of zero oxygen calibration solution (product ZERO OXYSOLN available from WAVE). Dip the bare probe in to the liquid. Wait for the reading to stabilize, then from the DOOPT screen: Set the temperature adjustment to the ambient temperature: o Press T/COMP. Using the pop-up keypad enter the ambient temperature. Press ENT to accept the new value.

Next push the CALIB button. Push the Set 0% button. A popup window will display SETTING 0%. Once the adjustment is complete the popup window will vanish.

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This sets the 0% oxygen point. Continue on to set the 100% point. Pure nitrogen gas (very low pressure) can be used in place of the zero oxygen solution.

2. 100% air Saturation: CALIBRATION FAIL If unit is not able to set the 0 or 100% points, a CAL FAIL message will appear. Retry the cal procedure and replace the probe and repeat. Remove the probe from the zero oxygen solution. Rinse the probe in water and shake dry. DO NOT TOUCH THE PROBE TIP OR ATTEMPT TO DRY IT WITH A CLOTH. Allow the probe to equilibrate in air. When the reading is stable in 1-2 minutes: Press CALIB again. Now press the Set 100% button. A popup window will display SETTING 100%. Once the adjustment is complete, the popup window will disapear.

This procedure sets the 100% oxygen point and completes the dissolved oxygen adjustment sequence.

REMEMBER TO SET THE TEMPERATURE COMPENSATION READING TO THE BIOREACTOR OPERATING TEMPERATURE.

Installation in Bioreactor
PREPARE,THE OXYWELLTM
Insert this end of the DOOPT probe into Oxywell2 fitting Locate the Oxywell2 fitting on the Cellbag. Remove the luer cap from the fitting and discard. Remember the Oxywell2 tip inside the Cellbag is sealed so removing this cap does not impact sterility. Fill the Oxywells with distilled water using the fill kit provided with the DOOPTPROBE. Instructions are included with the probe.

INSTALL THE DOOPT PROBE

After adjusting the DOOPT-PROBE, insert the probe carefully into the Oxywell2. Some older Cellbags have the original Oxywell fitting. This has a twin blue/white Y-connector. DOOPT cannot be used with this fitting. Please contact WAVE to get Cellbags with the Oxywell2 fitting. Secure the DOOPT-PROBE by tightening the luer connector. This also ensures that the probe is correctly seated in the fitting. The unique Oxywell2 sheath is an oxygen permeable diptube. This allows the DOOPT-PROBE to be removed and reinserted repeatedly without any danger of contamination. The Oxywell2 is completely sealed and allows only oxygen entry by diffusion through the silicone rubber tip.

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FOR OPTIMUM ACCURACY readjust the probe after insertion into the bag. This is possible if the Cellbag has not been inoculated. With aeration and rocking, this should correspond to the 100% saturation point. Let the Cellbag equilibrate so that it is at the correct temperature and at 100% air saturation. SET THE MANUAL TEMPERATURE COMPENSATION TO THE TEMPERATURE OF THE CELLBAG AND READJUST THE 100% POINT AS FOLLOWS: Set the temperature adjustment to the bioreactor temperature: o o Press the displayed temperature compensation value. Using the pop-up keypad enter the bioreactor operating temperature. Press ENT to accept the new value.

Press CALIB, now press the Set 100% button. A popup window will display SETTING 100%. Once the adjustment is complete the popup window window will disappear.

It is critical to fill the Oxywell2 with distilled water to provide a response time (t90) of about 3-5 minutes (see Technical Reference section).

PROBE MAINTENANCE AND STORAGE

The DOOPT-PROBE can be reused for many months. However, due to photobleaching, the amplitude of the fluorescent reading will ultimately become too low for accurate measurement (< 2000). Increasing the sampling frequency will shorten the probe life. The DOOPT module monitors the signal amplitude automatically, and will warn the user when the probe should be replaced. Unlike polarographic probes that are prone to sudden catastrophic failure, the DOOPT- PROBE performance decays very slowly and provides ample warning before the probe becomes unusable. No polarization or warm-up time is necessary. Unlike electrochemical probes, the optical DOOPT-PROBE does not contain any buffer to dry out and does not need to be stored wet or refrigerated. The DOOPT-PROBE fiber tip and coating can be easily damaged. Avoid hitting the probe on any hard surface or prying or scraping the fiber tip. The white fiber optic cable is quite flexible, but excessive force or a severe bend can snap the cable.

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EXTENDING PROBE LIFE

Probe life depends on the number of cumulative oxygen readings that are taken. Reducing the frequency of measurement will increase life, conversely increasing it will shorten probe life. The sampling frequency is set by the factory to once every 10 seconds but can be changed by the user from 1 to 99 seconds: Press on the displayed SCAN time value and enter the new scan time using the pop-up keypad. Press ENT to accept the new value. The instrument will now operate at the new sampling frequency.

VIEWING PROBE PERFORMANCE

DETAIL: Pressing DETAIL brings up a pop-up window that display various probe parameters, such amplitude and phase of the signal. This display is useful for troubleshooting and is shown in Figure 33. Amplitude: This window indicates the signal strength of the DO probe. The pH probe degrades with use over time. A reading over 2000 in the display window indicates a fully functional probe. When the reading is below 2000, the probe should be replaced. Phase: Refer to the Technical Reference section of this manual for a complete description of phase angle. Temperature: Indicates the temperature of the DO probe in the cellbag. Error Code: Refer to the Troubleshooting section of this manual for specific error descriptions in the Optical Dissolved Oxygen Error Codes Table.

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DO CONTROLLER PARAMETERS
DO controller parameters can be set from the SETUP menu -> DO. Access is restricted by password. Password 2050. DO parameter screen:

Figure 34. DO Control Setup Screen.

Setting Deviation Alarms:

DEV Alarm: If this parameter is set to OFF then deviations from the setpoint will not be monitored. Default: On. DEV Limit: Set the DO deviation alarm deadband. A deviation alarm is triggered if the DO is different from the DO setpoint by +/- this amount. The deviation alarm is only active if DEV alarm is ON and the DO controller is ON. Range 0.0 to 10%, Default: 10 %sat.

DO controller options:
DBAND (Deadband): Control action takes place if the DO is within the deadband. The deadband is defined +/- the setpoint. Increasing the setpoint will result in coarser control. Making the deadband too narrow will result in excessive chatter and poor control. Range 0.0 to 10%, Default: 5 % sat. CTIME: After each control action, the controller must wait for a defined cycle time (CTIME) in order to let the change result in a stable DO reading. If the CTIME is set too short then the controller will overshoot. If it is set too long, then the DO may not be controlled properly and drift. The CTIME can be set by the user to match process requirements and acid/base strength. Range 10 -999 seconds, Default: 300 seconds.

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Available Control Schemes

Two control schemes are defined. Either can be selected by the user: SPEED: The controller responds to change in DO outside the deadband by changing the BASE rocking speed setpoint. This option requires that the BASE be connected and configured in the WAVEPOD. A low DO results in a increase in speed. A high DO results in a decrease in speed. The speed is adjusted by STEP rpm every (cycle time in seconds) to give the DO sufficient time to equilibrate. The maximum and minimum rocker speed limits can be set by the user. An alarm is set when either of these limits are reached. STEP: This is the amount by which the controller changes the rocker speed setpoint at each cycle. Range 0.0 to 10 rpm, Default: 2 rpm. MINRPM: The controller does not reduce the BASE speed setpoint below this limit. Range 2 to 40 rpm, Default: 5 rpm. MAXRPM: The controller does not increase the BASE speed setpoint above this limit. Range 2 to 40 rpm, Default: 40 rpm. NOTE: Extremely low speed and low angle settings on the BASE are not recommended. O2 Concentration Control: The controller responds to change in DO outside the deadband by changing the O2% setpoint. This option requires that the O2 control module be installed, supplied with O2 gas and be turned on. The AIRPUMP must also be ON. A low DO results in an increase in O2 concentration. The concentration is adjusted by O2STEP every cycle time in seconds to give the headspace sufficient time to equilibrate. The maximum and minimum O2% concentration in the gas supplied to the headspace can be set by the user. An alarm is set when either of these limits are reached. STEP (%O2): This is the amount by which the controller changes the CO2 controller setpoint at each cycle. Range 0.0 to 1.0%O2, Default: 1% O2. MIN %O2: The controller does not reduce the CO2 setpoint below this limit. Range 0.0 to 50.0% O2, Default: 21 % O2. MAX %O2: The controller does not increase the CO2 setpoint above this limit. Range 0.0 to 50.0% O2, Default: 50 % O2.

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Oxygen Control
FUNCTION
The O2 module is used to inflate and ventilate the Cellbag disposable bioreactors with air containing a variable O2 concentration. O2 from a pipeline or cylinder is connected to the O2 module. Air is drawn from the AIR IN port (airpump module) and mixed with the O2 (connected to O2 IN). An O2 sensor measures the O2 concentration and controls it to any user-specified value from 21 to 50%. The O2 conditioned air is pumped from the MIX OUT port into the headspace of the Cellbag to maintain bag pressure, and to provide oxygen. If an oxygen supply is not available, consider using the O2GEN oxygen generator available from WAVE.

Vented air
MIX OUT AIR IN

Air

AIRPUMP

O2 Gas supply

O2 IN

Cellbag bioreactor

O2 module

O2 MODULE

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OPERATION
NOTE: A GREEN indicator will be displayed on the O2 CONTROL screen if the O2 inlet pressure is sufficient for operation. A red indicator shows that the O2 pressure is too low for proper control. 1. Connect power to the instrument. Switch ON. Allow 5 minutes for the unit to warm up and stabilize. 2. Set the external O2 source (line or O2GEN) at max pressure of 0.7- 1.0 bar (10-15 psig). Connect it to the O2 IN inlet connector located on the side. The unit is equipped with an internal non-adjustable pressure regulator. 3. Leave the AIR IN port unconnected if you wish to mix ambient air with the O2 stream. You can connect process air to this inlet. External gas pressure MUST be regulated to between 0.1 and 0.2 bar (1 3 psig). 4. The O2/air mixture is pumped out from the MIX OUT port on the front panel. Connect this to the AIR IN PORT on the 2050EHT BASE unit. For standalone operation, the MIX OUT can be connnected directly to the inlet filter on the Cellbag. 1. Air with up to 50% O2 content can be produced by the O2 module. Controlling the O2 content is an effective means of improving oxygenation. The O2 uses a zirconium oxide O2 sensor to measure the actual O2 content of the gas mixture. A feedback controller and valve is used to regulate the O2 content to the desired value. Since the actual oxygen concentration in the mix is measured and controlled, it is not necessary to feed 100% oxygen to the O2IN port. Any oxygen containing gas can be used as the controller will draw in as much of this gas as needed to produce the desired mix. CAUTION: If the pressure at the MIX OUT port exceeds 3 inches water gauge due to blockage or obstruction of the air flow to the Cellbag, a red high pressure LED indicator starts blinking on the O2CONTROL screen. If the overpressure condition continues for longer than 1-2 minutes, the internal air flow valve shuts down preventing the pressure from rising higher . When the overpressure condition clears, the airpump resumes operation.

NOTE: If you wish to produce an oxygen mixture with concentrations below 21% (air), you can connect a nitrogen stream to the AIR IN port.

NOTE: High pressure alarm does not activate if the airpump is in remote operation.

ADJUSTMENT
AERATION PUMP
1. Connect a certified flow meter to MIX OUT. 2. Turn on the Airflow. Set the setpoint to 0.5 lpm. 3. Go to the O2MIX Screen. 4. Compare the flow reading on the flow meter to AIR PV on the CONTROL SETUP screen. The readings should be within +/- 0.1 lpm.

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5. If the flow reading on the flow meter is higher than the AIR SP, increase the SCALING value. If the flow reading on the flow meter is lower, decrease the SCALING value. 6. Check the flow rate at different setpoints. 7. If the flow readings are not linear, contact WAVE for service.

O2 SENSOR
The O2 sensor does not need to be adjusted. Over time (10 years typical) the sensor will decay and will need to be replaced. Adjustment can easily be checked by following this procedure: 1. Allow the unit to warm up 5 minutes before adjusting. 2. Verify that the O2 gas is NOT connected to O2 IN and the O2 control is turned off. 3. Turn on the Airflow. Set the setpoint to 0.5 lpm. 4. Go to the O2 Control Setup Screen. Make sure that the O2 Offset is set to 0.0%. 5. Verify that the O2 PV display reads 21.0% (+/- 0.2%). If the reading is not within tolerance, adjust the O2 offset. 6. Connect 45% Certified O2 gas to AIR IN. Set the flow to 0.5 1.0 lpm. 7. Verify that the O2 PV display reads 45.0% (+/- 0.2%). If the O2 reading is not linear, replace the sensor. For sensor replacement, please contact WAVE for return of the unit for service.

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O2 CONTROL SCREEN
Pressing the O2MIX button (OVERVIEW screen) brings up the O2MIX CONTROL screen:

Figure 35. O2MIX Control Remote Base Screen. IMPORTANT: O2 gas must be connected to the O2 IN port on the side of the unit. The pressure must be regulated to between 10-15 psig (0.7 to 1.0) bar and capable of delivering at least 0.2 lpm. O2 PV (Process Variable): The actual O2 concentration in %O2 is shown in the box labeled %O2. O2 Setpoint: The %O2 setpoint is shown in the box labeled SP. Pressing the setpoint brings up a data entry keypad. You can enter a new setpoint and press the ENT key to accept the new setpoint. The keypad does not permit data entry outside a preset range. Setpoint Range is 0 to 50% O2, Control Range is 21 to 40% +/- 0.6%, 41 to 50% +/- 1.0%. O2 ON/OFF: Pressing this button starts O2 control. The button changes to green and displays ON. Pressing it again disables O2 control and the button changes to red and displays OFF. RSP (Remote set point): The O2 setpoint can be set remotely by another controller. Typically, the dissolved oxygen (DO) controller can be selected to remotely change the O2 in order to maintain a set dissolved oxygen concentration in the bioreactor. When the option is active, a flashing message -> RSP from DO will be displayed. Trends: The O2 is graphed over a 10 minute period. Press the TREND button to view the graph. Air flow is required for the O2 controller to work properly. For convenience, airflow controls are provided on the right side of the O2 screen: MIX OUT Airflow: The actual airflow in liters/minute is shown in the box labeled lpm. Airflow Setpoint: The airflow setpoint is shown in the box labeled SP. Pressing the setpoint brings up a data entry keypad. You can enter a new setpoint and press the ENT key to accept the new setpoint. The keypad does not permit data entry outside a preset range. Range is 0 to 0.5 lpm. AIR ON/OFF: Pressing this button starts airflow control. The button changes to green and displays ON. Pressing it again disables airflow control and the button changes to red and displays OFF (LOCAL MODE ONLY).

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Trends: The airflow is graphed over a 10 minute period. Press the TREND button to view the graph.

O2 MIX CONTROL PARAMETERS SCREEN

O2 concentration controller parameters can be set from the SETUP menu -> O2. Access is restricted by password.

Figure 36. O2 Mix Control Setup Screen.

Setting O2 Deviation Alarms:

DEV Alarm: If this parameter is set to OFF, then deviations from the setpoint will not be monitored. Default: On. DEV Limit: Set the O2 deviation alarm deadband. A deviation alarm is triggered if the O2 concentration is different from the O2 setpoint by +/- this amount. The deviation alarm is only active if DEV alarm is ON and the O2 controller is ON. Range: 0.0 to 5.0%, Default: 2 %.

O2 controller adjustment:
O2OFFSET: Small adjustments can be made by changing the offset. See ADJUSTMENT section for proper usage. Default: set at factory.

Setting AIRFLOW Deviation Alarms:

DEV Alarm: If this parameter is set to OFF, then deviations from the setpoint are not monitored. Deviation limit is fixed at 0.02 lpm. Default: On.

AIRFLOW controller adjustment:

SCALING (ml/min): Small airflow adjustments can be made by changing the scaling. See ADJUSTMENT section for proper usage. Default: set at factory.

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CO2 Control
FUNCTION
The CO2 module is used to inflate and ventilate the Cellbag disposable bioreactors with air containing a variable CO2 concentration. CO2 from a pipeline or cylinder is connected to the CO2 module. Air is drawn from the AIR IN port by the airpump and mixed with the CO2 (connected to GAS IN). A CO2 sensor measures the CO2 concentration and controls it to any user-specified value from 0 to 15%. The CO2 conditioned air is pumped from the MIX OUT port into the headspace of the Cellbag to maintain bag pressure, and to provide oxygen and carbon dioxide for ventilation and pH control.

Vented air
MIX OUT AIR IN

Air

AIRPUMP

CO2 Gas supply

CO2 IN

Cellbag bioreactor

CO2 module

CO2 MODULE

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CO2 OPERATION
1. Connect power to the instrument. Switch ON. Allow 5 minutes for the unit to warm up. NOTE: A GREEN indicator will be displayed on the CO2 CONTROL screen if the CO2 inlet pressure is sufficient for operation. A red indicator shows that the CO2 pressure is too low for proper control. 2. Set the external CO2 source (line or cylinder) to between 0.7 and 1 bar (10-15 psig). Connect it to the CO2 IN inlet connector located on the side of the unit. The unit is equipped with an internal pressure regulator requiring no user adjustment. 3. Leave the AIR IN port unconnected if you wish to mix in ambient air with the CO2 stream. Alternatively, process air can be connected. Process gas pressure MUST be regulated to between 0.1 and 0.2 bar (1- 3 psig). 4. The CO2/air mixture is pumped out from the MIX OUT port on the side panel. Connect this to the AIR IN PORT on the 2050EHT BASE unit. For standalone operation, the MIX OUT can be connnected directly to the inlet filter on the Cellbag. Air with up to 15% CO2 content can be produced by the CO2 module. Controlling the CO2 content is an effective means for pH control using media containing bicarbonate buffers. Increasing the CO2 content lowers the pH. The CO2 module uses a NDIR CO2 sensor to measure the actual CO2 content of the gas mixture. A feedback controller and valve is used to regulate the CO2 content to the desired value. NOTE: High pressure alarm does not activate if airpump is in remote operation. CAUTION: If the pressure at the MIX OUT port exceeds 3 inches water gauge due to blockage or obstruction of the air flow to the Cellbag, the red high pressure LED will start blinking. If the overpressure condition continues for longer than 1-2 minutes, the integral air flow valve will shut down preventing the pressure from rising higher. When the overpressure condition clears the airpump will resume operation.

AIRFLOW CONTROLLER ADJUSTMENT:

SCALING: Small airflow adjustments can be made by changing the scaling. See ADJUSTMENT section for proper usage. Default: set at factory.

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CO2 CONTROL SCREEN

Pressing the CO2 button (OVERVIEW screen) brings up the CO2 MIX CONTROL screen:

IMPORTANT 100% CO2 gas must be connected to the CO2 IN port on the side of the unit. The pressure must be regulated to between 10-15 psig (0.7 to 1.0) bar and capable of delivering at least 0.2 lpm

Figure 37. CO2Mix Control Remote Base Screen. CO2 PV (Process Variable): The actual CO2 concentration in %CO2 is shown in the box labeled %CO2. CO2 Setpoint: The %CO2 setpoint is shown in the black SP box. Pressing the setpoint brings up a data entry keypad. The keypad does not permit data entry outside a preset range. Range is 0 to 15% CO2 CO2 ON/OFF: Pressing this button starts the CO2 control. The button changes to green and displays OFF. Pressing it again disables CO2 control and the button changes to red and displays OFF. RSP (Remote set point): The CO2 setpoint can be set remotely by another controller. Typically, the pH controller can be selected to remotely change the CO2 in order to maintain pH. When the option is active, a flashing message -> RSP from pH is displayed. Trends: The CO2 is graphed over a 10 minute period. Press the TREND button to view the graph. Air flow is required for the CO2 controller to work properly. For convenience, airflow controls are provided on the right side of the CO2 screen: MIX OUT Airflow: The actual airflow in liters/minute is shown in the box labeled lpm. MIX OUT Airflow Setpoint: The airflow setpoint is shown in the box labeled SP. Pressing the setpoint will bring up a data entry keypad. You can enter a new setpoint and press the ENT key to accept the new setpoint. The keypad does not permit data outside a preset range. Range is 0 to 1.00 lpm. AIR ON/OFF: Pressing this button starts airflow control. The button changes to green and displays ON. Pressing it again disables the airflow control and the button changes to red and displays OFF (LOCAL MODE ONLY).

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46

Trends: The airflow is graphed over a 10 minute period. Press the TREND button to view the graph.

CO2MIX CONTROL SETUP

CO2 concentration controller parameters can be set from the SETUP menu -> CO2. Access is restricted by password. Password 2050.

CO2 PARAMETER SCREEN:

Figure 38. CO2 Mix Control Setup Screen.

SETTING CO2 DEVIATION ALARMS:

DEV Alarm: If this parameter is set to OFF then deviations from the setpoint will not be monitored. Default: On. DEV Limit: Set the CO2 deviation alarm deadband. A deviation alarm is triggered if the CO2 is different from the CO2 setpoint by +/- this amount. The deviation alarm is only active if DEV alarm is ON and the CO2 controller is ON. Range: 0.0 to 5.0%, Default: 1 %.

CO2 CONTROLLER ADJUSTMENT:

CO2OFFSET: Small adjustments can be made by changing the offset. See ADJUSTMENT section for proper usage. Default: set at factory.

SETTING AIRFLOW DEVIATION ALARMS:

DEV Alarm:. If this parameter is set to OFF then deviations from the setpoint will not be monitored. Deviation limit is fixed at 0.02 lpm. Default: On.

AIRFLOW CONTROLLER ADJUSTMENT:

SCALING: Small airflow adjustments can be made by changing the scaling. See ADJUSTMENT section for proper usage. Default: set at factory.

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ADJUSTMENT
AERATION PUMP
1. Connect a certified flow meter to MIX OUT. 2. Turn on the Airflow. Set the setpoint to 0.5 lpm. 3. Go to the CO2MIX CONTROL SETUP screen 4. Compare the flow reading on the flow meter to AIR PV on the O2MIX CONTROL SETUP or CO2MIX CONTROL SETUP screen. The readings should be within +/- 0.2 lpm. 5. If the flow reading on the flow meter is higher than the air AIR SP, increase the SCALING value. If the flow reading on the flow meter is lower, decrease the SCALING value. 6. Check the flow rate at different setpoints. 7. If the flow readings are not linear, contact WAVE for service.

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Maintenance

Routine Maintenance
Cleaning
The exterior of the WAVEPOD should be cleaned with a damp cloth with water. Clean the unit on regular basis and exam all tubing connections daily. DO NOT use abrasive cleaners.

Fuses
Two fuses are located inside the power entry module. Fuse Type, 3A, 250V, 5 x 20mm, slow-blow To replace the fuses: CAUTION: Disconnect power cord before attempting to replace the fuses. 1. Disconnect power and detach the power entry cable. 2. Locate the power entry module and pry off the fuse cover. 3. Remove and replace the fuses. 4. Replace the module cover. 5. Reattach power cord and power up.

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CO2 Sensor Adjustment

The CO2 sensor needs to be readjusted every six months to ensure accuracy. This may be done by returning the unit to the WAVE, or by following this procedure:

CO2 OFFSET ADJUSTMENT

1. Turn power off. Loosen the four panel screws on the CO2 module, slide out the module. Disconnect the tubing to the CO2 sensor. Install the extender card and connect the CO2 module. Extender cards are available from WAVE, contact the service department. Make sure to support the module so it remains connected in the extender card. 2. Turn power on. Allow the unit to warm up 5 minutes before adjusting. 3. Go to the CO2 Control Setup Screen (Press SETUP, enter 2050, CO2). Verify that the CO2 Offset is set to 0.0%. 4. Connect 100% nitrogen gas the CO2 sensor. Set the nitrogen flow between 0.5 1.0 lpm. 5. Verify that the CO2 display is reading 0.0% (+/- 0.25%). Allow the reading to stabilize. 6. Connect certified 10 % CO2 gas to the CO2 sensor. Set the CO2 flow between 0.5 1.0 lpm. 7. Verify that the CO2 display is reading 10.0% (+/- 0.25%). Allow the reading to stabilize. If the CO2 reading is not within tolerance, adjust the CO2 offset accordingly. 8. Re-check 0.0%. Connect certified 5.0% CO2 to the CO2 sensor. Set the flow between 0.5 1.0 lpm. Allow the reading to stabilize. Verify that the CO2 reading is 5.0% (+/- 0.25%). 9. The CO2 readings should be linear at 0%, 5% and 10% (+/- 0.25%). 10. Repeat Steps 4 8 if necessary. 11. If the CO2 readings are not linear then CO2 Sensor Board adjustment is necessary.

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CO2 SENSOR BOARD ADJUSTMENT

1. Make sure that the CO2 offset is set to 0.0. 2. Turn power off to the unit, then, loosen the four panel mount screws. 3. Remove the CO2 sensor module from the rack, disconnect the tubing. Slide in an extender card for the module. 4. Connect the CO2 sensor module to the extender card. Make sure to support the module. 5. Turn power on. Allow 5 minutes for the instrument to warm up. 6. Connect the BLACK (-) lead of a multimeter to JP1 (Signal Common) and the RED (+) lead to TP7 (0 1 V Output). 7. Verify that the CO2 sensor is set to the proper control range. Press and Hold SW4. Verify that the multimeter reads 1.00V dc. If it does not read 1.00 V dc, press and hold SW4. Use SW1 as an UP and SW3 as a down switch to adjust the voltage reading. 8. Verify that the CO2 sensor is set to the proper adjustment gas value. Press and Hold SW2. Verify that the multimeter reads 0.50 V dc. If it does not read 0.50 V dc, press and hold SW2. Use SW1 as an UP and SW3 as a down switch to adjust the voltage reading. 9. Connect 100% nitrogen gas to CO2 sensor. Set the nitrogen flow between 0.5 1.0 lpm. Allow the gas to flow for at least one minute. 10. Press and hold SW1 for 2 seconds. The volt reading on the multimeter should read 0.0 V dc (+/- 0.01). Do not allow the volt reading to go negative. 11. Disconnect the nitrogen gas and connect 10% Certified CO2 gas to the CO2 sensor. Set the CO2 flow between 0.5 1.0 lpm. Allow the gas to flow for at least one minute. 12. Press and hold SW3 for 2 seconds. The volt reading on the multimeter should read 0.50 V dc (+/- 0.01). 13. Go to the CO2 Control screen, verify that the CO2 display reads 10.0% (+/-0.25%). If the reading is not within tolerance, adjust VR1(Coarse adjustment) or VR2 (Fine Adjustment). 14. Reconnect 100% nitrogen, Allow the gas to flow for at least one minute. Verify that the CO2 reading is 0.0% (+/- 0.25%). 15. Disconnect the nitrogen gas and connect 5% Certified CO2 to AIR IN. Set the flow to 0.5 1.0 lpm. Allow the gas to flow for at least one minute. 16. Verify that CO2 display reads 5.0% (+/- 0.25%). 17. If the CO2 readings are not linear, repeat steps 9 - 16. If the CO2 readings are still not linear, replace the CO2 sensor. 18. Disconnect the gas, turn power off. Remove the extender card, connect the tubing and install the module.

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For sensor replacement contact please contact WAVE for return of unit for service.

Replacement Module Parts List

The WAVEPOD modules can be ordered individually and can typically be installed by the user. The following table lists the modules and their part numbers. Module Description Bay Location 1 2 3 4 4 4 pH Control Module Dissolved Oxygen Control Module Airpump Module O2 Gas Concentration Control Module CO2 Gas Concentration Control Module Combination O2 + CO2 Control Module WAVE Part Number POD.003607 POD.003605 POD.003601 POD.003603 POD.003604 POD.003602

NOTE: Bay number 4 can accommodate either: O2, CO2 or combination O2 + CO2 modules.

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Troubleshooting
GENERAL PROBLEMS
Display stays blank
The touchpanel should display the startup screen a few seconds after power up. If the startup screen does not appear, verify that you have power by checking the Ethernet status LED. If it is not lit then check the unit fuse and power cord.

Keep getting BASE NOT FOUND MESSAGE

This message is displayed momentarily on startup and if communications to the connected base are lost. If the message stays on then internal communications to WAVEPOD control modules have been lost. Contact WAVE service for assistance.

OPTICAL DISSOLVED OXYGEN

Low amplitude alarm Check that the DOOPT-PROBE is connected correctly
and ensure that the plug is secure. Cycle power to the instrument to reset. If warning persists, replace the DOOPT-PROBE. Check the DETAIL display for the signal amplitude. Less than 2000 indicate a bad probe.

Fluctuating reading Some fluctuation (+/- 5%), especially at high DO levels

is normal.

Probe adjustment is drifting The DOOPT-PROBE should be readjusted

every 2-4 days to compensate for drift. If the drift is greater than 10%, the probe should be replaced. The following table lists the error codes for the Optical Dissolved Oxygen Probe. ERROR CODE 0 1 4 DESCRIPTION ADC overflow High ambient light Low amplitude

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CO2 MIX
Display shows 0.1 even in air The tolerance of the CO2 sensor is from 07.5%, +/- 0.38% CO2, so this reading is not unusual. If zero is suspect, then readjust the sensor as previously described.

CO2 reading keeps drifting down The CO2 gas supply pressure is too low. Check that the green CO2 inlet pressure light is lit. The CO2 supply pressure must between 10 and 15 psig. CO2 is not controlling properly Verify that CO2 is connected to the correct
inlet port. Check that the Air pump is on and that the flowrate is between 0.1 and 0.5 liters/minute. Check that the CO2 switch is ON. Check the setpoint on the controller.

No airflow reading Check that airflow to Cellbag is not obstructed. Verify

that the pump is working by disconnecting the MIX OUT coupling on the instrument.

Cellbag does not stay inflated Verify that there is airflow to the Cellbag. Check that the check valve on the Cellbag is correctly installed. Close off the Cellbag outlet using the pinch clamp and see if the Cellbag inflates. Then confirm that the check valve is working by connecting a short length of tubing to the check valve outlet and dip it into water. Tubing should be immersed to a depth of 5-10 mm. Opening the outlet pinch valve should cause bubbles to appear and bubbles should continue to flow as the flow and pressure stabilizes.

O2 MIX
Display shows 21.4% O2 even in air The tolerance of the O2 sensor is from 21 to 40% O2, +/- 0.6%, so this reading is not unusual. If zero is suspect, then readjust the sensor as previously described. O2 reading keeps drifting down The O2 gas supply pressure is too low.
Check that the green O2 inlet pressure light is lit. The O2 supply pressure must between 10 and 15 psig.

O2 is not controlling properly Verify that O2 is connected to the correct inlet port. Check that the O2 supply pressure is set to 0.7 bar minimum. Check that the Air pump is on and that the flowrate is between 0.1 and 0.5 liters/minute. Check that the O2 switch is ON. Check the setpoint on the controller. No airflow reading Check that airflow to the Cellbag is not obstructed.
Verify that pump is working by disconnecting the MIX OUT coupling on the instrument.

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ALARM MESSAGES
Alarm # 01 02 03 10 11 12 20 21 30 31 32 40 41 42 43 44 45 46 47 48 49 50 51 Message BAY1&3 FAIL BAY2&4 FAIL BASE COM FAIL PH_DEV PH_FAIL PHCOM_FAIL DO_DEV DO_FAIL INIT_FAIL AIR_DEV OUTHIPRALM O2_DEV CO2_DEV O2_FAIL CO2_FAIL LOWO2PR LOWCO2PR O2MINLMT O2MAXLMT CO2MAXLMT CO2MINLMT RPMMINLMT RPMMAXLMT Action Communications with BAY1&3 lost Communications with BAY2&4 lost Communications with BASE unit lost pH deviates from setpoint pH sensor failed or disconnected pH amplifier not responding DO deviates from setpoint DO sensor failed or disconnected PLC initialization failure Airflow deviates from setpoint MIX OUT pressure too high O2 deviates from setpoint CO2 deviates from setpoint O2 sensor failed CO2 sensor failed Oxygen supply pressure low CO2 supply pressure low O2 remote setpoint at minimum limit O2 remote setpoint at maximum limit CO2 remote setpoint at maximum limit CO2 remote setpoint at minimum limit speed remote setpoint at minimum limit speed remote setpoint at maximum limit

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BASE ALARM MESSAGES 2050EHT

Alarm # E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 E13 E14 E15 E16 E17 E18 E19 E20 E21 E22 E23 E24 E25 E26 E27 E28 E29 E30 Message INIT FAIL SPEED DEV AIR DEV TEMP DEV WEIGHT DEV TEMPFAIL HTR FAIL ROCKFAIL COMMFAIL MTRFAIL HIPRALM_L EMERSTOP CO2FAIL PANTFAIL RTEMPFAIL RTEMPDEV RAIRDEV CO2DEV HIPRALM_R LOADFAIL LOWWTALM HIWTALM HARVALM FEEDALM LOCO2PR CODE_FAIL O2FAIL LOO2PR O2DEV LOAD_OVRG Action Machine failed to initialize on power up Actual rocking speed does not match setpoint Actual airflow (left) does not match setpoint Actual temperature (left) does not match setpoint Actual weight does not match setpoint Temperature sensor (left) failed or unplugged Heater unplugged or over temperature Rocker not rocking Internal communications failure. Cycle power to retry Rocking motor failure High bag (left) pressure. Safety bumper activated and stopped rocker CO2 sensor fail (2050EHT-CO2 only) Not used for bioreactor Right side temperature sensor failed or unplugged (2050EHTD) Actual temperature (right) does not match setpoint (2050EHTD) Actual airflow (right) does not match setpoint (2050EHTD) CO2 deviation (2050EHT-CO2 only) High bag (right) pressure (2050EHTD only) Weight sensor failed (only when loadcell is installed) Low weight alarm (only when loadcell is installed) High weight alarm (only when installed) loadcell is Harvest alarm (only when loadcell is installed) Feed alarm (only when loadcell is installed) Low CO2 supply pressure (2050EHT-CO2 only) Bar code reader failed O2 sensor fail (2050EHT-O2 only) Low O2 supply pressure (2050EHT-O2 only) O2 deviation (2050EHT-O2 only) Weight exceeds sensor range

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Communications & Data Acquisition

OVERVIEW
Two options for data acquisition are built into the WAVEPOD: 1. ALARM: Currently, a unit alarm is available on the back panel jack. This is a normally closed (or NO) contact that will open if a unit alarm occurs or power fails. This is a dry contact output that can be used to connect to user alarm monitoring systems. 2. DIGITAL: Digital communication ports are provided on the back panel for RS-485 connection of a Wave Bioreactor BASE unit, such as a SYSTEM20/50EHT, 20/50EH or 02/10EH.). A 10BaseT Ethernet port is provided for data acquisition and supervisory control

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ALARM PORT
A DB15 female jack on the back panel provides the alarm contacts. Use a standard DB15 male plug to connect to this port. The port pinout is listed in the table below. Pins 1 through 10 are reserved for future use. All signals are protected against short circuit. The default pinouts are shown in the table below: PIN 1-10 11 12 13 14 15 DEFAULT PARAMETER Reserved for future use. Factory use only Unit Alarm + contact closes on alarm Unit Alarm - contact closes on alarm Unit Alarm + contact opens on alarm Unit Alarm - contact opens on alarm Dry contact Dry contact Dry contact Dry contact Contact

ALARM CONTACT
A unit alarm is provided. This dry contact can be wired open-on-alarm (failsafe). In this configuration, the contact will open if an alarm occurs or if power is lost to the instrument. If the close-on-contact pins are used, then the logic works in the opposite fashion and contact will close on alarm. No power loss indication is possible in the latter configuration. Alarm contacts are rated at a load voltage 350V (Peak) and at a load current of 120mA (continuous). The unit alarm is tripped on various alarm conditions as configured using the touchscreen. See Alarm Messages. Alarm limits can be changed from the SETUP menus. Individual alarms can also be enabled and disabled.

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BASE COMMUNICATION PORT

A RJ12-6 telephone style jack is provided on the rear to connect to a Wave Bioreactor BASE unit. Use a RJ12- RJ12 straight cable (WAVE part # 001833) to connect the WAVEPOD to the appropriate base. The cable should plug into one of the two DATAPORT jacks on the BASE unit. DO NOT PLUG ANYTHING INTO THE UNUSED DATAPORT ON THE BASE UNIT. Use shielded cable and cable length should not exceed 15 feet. The communications parameters are: 9600 baud No Parity 8 data bits 1 stop bit The communication protocol used is standard MODBUS RTU. 1 RJ11 RS-485 MODBUS JACK PINOUT RJ11 pin 2 RJ11 pin 5 RJ11 pin 4 DATA A(-) DATA B(+) SIGNAL GND RJ11 jack pinout viewed looking at jack 6

BASE-POD INTERCONNECT CABLE

Connect to a DATAPORT jack on BASE unit

Connect to BASE jack on WAVEPOD

RJ11 6 conductor Style II straight cable

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PROGRAMMING PORTS
NOTE: Key must be in RUN POSITION to operate the WAVEPOD. Set to PROG for programming only. A DB9 jack (labeled PROG) is provided on the rear panel for programming. A special cable is required. To prevent unauthorized programming, the rear keylock MUST be in the PROG position. A key is provided with the WAVEPOD unit. A COMM port is provided for external communications such as printers. Consult WAVE for COMM port functionality.

ACID/BASE INTERFACE CABLE

Two DB9 jacks are provided on the rear panel for connecting PUMP20 acid and base pumps. These jacks provide power and control connections. ACID/BASE PINOUT 1 6 9 5 COMMON +24V power to pump dry contact terminal 1 dry contact terminal 2 PUMP1 PUMP2 5 9 acid pump base pump Use DB9-DB9 cable supplied with PUMP20 PUMP20

The dry contact will close (N/O) or open (N/C) when unit activates pump signal.

6 Wire remote contact to pin 9 and pin 5. Maximum rating 150mA.

User pump remote start/stop contact

CAUTION
DO NOT SWITCH AC VOLTAGE THROUGH DRY CONTACTS. DAMAGE TO THE UNIT WILL OCCUR.

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ETHERNET COMMUNICATIONS
For greater flexibility, WAVEPODs can be used to concentrate data from the associated bioreactor and make it available over Ethernet for data storage and supervisory control. Each WAVEPOD must have a unique IP address.

PCDAQ/SCADA PC server

Networked client PCs

Internet or corporate Ethernet Ethernet switch

MODBUS/Ethernet
192.168.0.1

WAVEPOD

Bioreactor

Ethernet
192.168.0.2

WAVEPOD
192.168.0.3

Bioreactor

Wireless access point link

WAVEPOD

Bioreactor

Converters are set at the factory to the IP address 198.162.0.1. This IP address can be changed by the user over Ethernet from a PC. Refer to the section in this manual Changing the IP address, see next page. Ethernet networks can be created using commercial Ethernet switches and hubs. Wireless and remote links can be configured. When using Ethernet, the communication protocol used is MODBUS Ethernet. To address any device you must give the driver the converter IP address and

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then the MODBUS address of the instrument. The data register numbers internal to the instrument are provided in Data Registers section. For example to read the actual rocking speed the driver will need: WAVEPOD IP address: Speed PV register: 192.168.0.1 42034 WAVEPOD MODBUS address: 1 (always)

ETHERNET PORT
A RJ45 Ethernet is provided on the rear panel to communication with external data acquisition and supervisory control programs. Standard procedures should be used to connect this port to a user network. Typically the WAVEPOD is connected to an Ethernet hub using a standard Cat5 Ethernet cable. Other devices may be connected to the network and PCs connected to the network can access data from the WAVEPOD.

NOTE: If two WAVEPODs are set to the same Ethernet address, the data acquisition program will not be able to uniquely identify them. You may get data from one or the other at random. Make sure all Ethernet addresses are unique. . NOTE: Changing the IP sometimes requires knowledge of the MAC address of the internal Ethernet converter. This is marked on the rear panel.

DIGITAL INSTRUMENT ADDRESSING

Each WAVEPOD can be addressed over the Ethernet port using its unique IP address. The default address is: 192.168.0.1. Other parameters are: Modbus address: Protocol: Notes: 1 (do not change) MODBUS Ethernet/TCP Maximum of 16 registers per MODBUS call.

Multiple WAVEPODs can be networked using standard Ethernet hubs and switches as described in the previous section. Each WAVEPOD must have unique IP address. The IP address of the built-in Ethernet interface can be changed using a PC running Lantronix configuration software (provided on CDROM with unit).

CHANGING THE IP ADDRESS

Use the following procedure to change the WAVEPOD IP address 1. Install the latest Lantronix configuration utility (www.lantronix.com>support-->downloads on a suitable PC. Select the Device Installer for UDS-10. Follow instructions to download and install. 2. Connect the PC to the WAVEPOD via Ethernet and verify correct link (LINK light on WAVEPOD Ethernet panel should turn green).

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3. Start the DeviceInstaller program. Click on the IP icon to assign new IP address. If the hardware address is blank then enter the hardware or MAC address manually. Click on Set IP to assign the new IP address. 4. Ping the new IP using DeviceInstaller. 5. Terminate DeviceInstaller and cycle power to the WAVEPOD to complete the procedure. It is helpful to label the new IP address on the unit.

ETHERNET DISPLAY STATUS

The front panel of the WAVEPOD contains an Ethernet display that provides information regarding link activity and diagnostics for troubleshooting. SERIAL LEDS 10 Mbps link/activity steady green 10 Mbps link/activity blinking 100 Mbps link/activity steady green 100 Mbps link/activity blinking Collision blinking red Diagnostic steady red and status blinking MEANING Valid 10 Mbps network connection Network packets transmitting and receiving Valid 100 Mbps network connection Network packets transmitting and receiving Network collisions 2 blinks = RAM 4 blinks = EEPROM checksum error 5 blinks = Duplicate IP address on network Diagnostic blinking red and status blinking green Status steady green Status blinking green 5 blinks = No DHCP response Serial port not connected to network Serial port connected to network

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Connecting to PCs Systems

Connecting the WAVEPOD to host PCs and supervisory host computer systems information: For users who are interested in developing their own data acquisition package, the WAVEPOD is based on an open architecture model. With a suitable MODBUS/TCP driver, the user can develop custom software. See Data Registers MODBUS memory mapping and register details.

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Data Registers
Data register information is useful for user-written external data acquisition programs. Only qualified personnel should attempt to access data registers as unpredictable operation may result. Data registers are 16 bit integers (range -32768 to +32767). Discrete I/O is bit of word. Scaling information is provided in the following tables:

EHT BASE Unit Dataset

MODBUS 42000 42001 42002 42003 42004 42005 42006 42007 42008 42009 42010 42011 42012 42013 42014 42015 42016 42017 42018 42019 42020 42021 42022 42023 42024 42025 42026 42027 42028 42029 42030 42031 Device EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT PV/SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP Description Command relay Speed SP Angle SP Airflow_L SP Temp_L SP Load SP Airflow_R SP (DUAL) Temp_R SP (DUAL) CO2 SP (CO2 option) O2 SP (O2 option) unused Feed rate/day Feed ml/shot Motor Accel SP unused Max Pan Temp SP unused unused unused unused unused unused unused unused unused unused unused unused unused unused unused unused Scaling rpmx10 degreex10 ml/min degCx10 net Kg/100 ml/min degCx10 CO2%x10 O2%x10

degCx10

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MODBUS 42032 42033 42034 42035 42036 42037 42038 42039 42040 42041 42042 42043 42044 42045 42046 42047 42048 42049 42050 42051 42052 42053 42054 42055 42056 42057 42058 42059 42060 42061 42062 42063

Device EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT EHT

PV/SP PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV

Description motor status angle PV speed PV airflow_L PV temp_L PV net weight R2 misc flags operating hours STATUS CONFIGURATION OPTIONS ALARM1 ALARM2 bag status 0 - 6 bag serial # x 1000 bag serial # - lower digits airflow_R PV temp_R PV CO2 PV cum feed cum harvest O2 PV

Scaling degreesx10 rpmx10 ml/min degCx10 gm/10 hours

ml/min degCx10 %CO2x10

%O2x10

TP communication check calc feed interval time to next feed

minutes minutes

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R/W

MOM

READ ONLY

READ ONLY

READ ONLY

READ ONLY

READ ONLY

42044

42043

42042

42041

42040

42000

MODBUS

SP registers can be written to: Set 1 momentarily to set. Program should clear after write (250 mS). PV registers are read only. For example, to check the speed deviation alarm, read MODBUS register 42043 bit 2.

WAVEPOD
SP TYPE PV 1 BIT PV PV PV PV

RAIR_DEV CO2_DEV HIPRALM_R LOAD_FAIL LOWWT_ALM HIWT_ALM HARVALM FEEDALM LOCO2PRALM BAR_FAIL O2_FAIL LOO2PRALM O2_DEV LOAD_OVRG PAN_TFAIL RTEMP_FAIL RTEMP_DEV CO2_FAIL EMERSTOP HIPRALM_L LRHTR_ENBL LOAD_ENBL CO2_DEVENBL DAC_ENBL BAR_ENBL INV_PUMP MTR_FAIL ROCKHTROFF COMM_FAIL NOHIPROFF ROCK_FAIL INTER_ROCK SNGL_HEAT HTR_FAIL BEEP_ENBL CO2_ENBL TEMP_FAIL AUTO_ACK PANHTR_EMBL RAIR_ON RHEAT_ON FEED_ON HARV_ON FEEDPMPON HARVPMP_ON PERHARV_ON O2_ON LOAD_DEV AUTOSTART DUAL_HEAT CO2_ON TEMP_DEV L_DEVENBL DUAL_AIR PERF_ON AIR_DEV T_DEVENBL O2_ENBL HEAT_ON HEAT_SW SPEED_DEV A_DEVENBL HEAT_ENBL LAIR_ON LAIR_SW

INIT_FAIL

S_DEVENBL

AIR_EMBL

ROCK_ON

ROCK_SW
2 3 4

PERF_SW
5

O2_SW
6

CO2_SW
7

Discrete I/O Table - EHT

RAIR_SW
8

RHEAT_SW
9

TARE_SW
10

FEED_SW HARV_SW RESET_CUM ACK_SW

11 12 13 14 15

67

16

WAVEPOD Dataset
MODBUS 42064 42065 42066 42067 42068 42069 42070 42071 42072 42073 42074 42075 42076 42077 42078 42079 42096 42097 42098 42099 42100 42101 42102 42103 42104 42105 42106 42107 42108 42109 42110 42111 Device POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 POD1 PV/SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV Description command relay Scaling

airflow SP pH SP

ml/min pHx100

TP communication check

pHOPT amplitude airflow PV R2 misc flags pH PV STATUS CONFIGURATION OPTIONS ALARM1 ALARM2 remote CO2 SP to POD2

ml/min pHx100

CO2%x10

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MODBUS 42128 42129 42130 42131 42132 42133 42134 42135 42136 42137 42138 42139 42140 42141 42142 42143 42160 42161 42162 42163 42164 42165 42166 42167 42168 42169 42170 42171 42172 42173 42174 42175

Device POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2 POD2

PV/SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP SP PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV PV

Description command relay O2SP CO2SP

Scaling O2%x10 CO2%x10

DOSP

DO%x100

Remote O2SP Remote CO2SP Remote RPMSP

O2%x10 CO2%x10 RPMx10

O2 PV

O2%x10

DOOPT amplitude CO2 PV R2 misc flags DO PV STATUS CONFIGURATION OPTIONS ALARM1 ALARM2

CO2%x10 DO%satx100

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Discrete I/O Table WAVEPOD POD1 (Bay 1&3)

BIT MODBUS 42064 R/W MOM TYPE SP 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

42104 READ ONLY

AIRPUMP_ON

PV

42105 READ ONLY

PV PHELECENBL PHOPTENBL AIR_ENBL

PV PH_DEVENBL PHCO2ENBL AUTOSTART AUTO_ACK

42107 READ ONLY

CO2MAXLMT

PV

42108 READ ONLY

PV

SP registers can be written to: Set 1 momentarily to set. Program should clear after write (250 mS). PV registers are read only. For example, to check the air deviation alarm, read MODBUS register 42107 bit 2.

CO2MINLMT

PHCOMFAIL

OUTHIPRALM

PH_FAIL

INIT_FAIL

AIR_DEV

PH_DEV

PhABENBL

READ ONLY

HIPRALM_ENBL

AIR_DDEVENBL

42106

PHENBL

ACDBAS_ON

PHCTRL_ON

PHCMD

WAVEPOD

SETPHSENS_SW 70

SETPHCAL_SW

SETPHSC_SW

SETPHTC_SW

PHCTRL_SW

SETPH7_SW

SETPH4_SW

AIRPMP_SW

R/W

MOM

READ ONLY

READ ONLY

READ ONLY

READ ONLY

READ ONLY

42172

42171

42170

42169

42168

42128

MODBUS

WAVEPOD
SP TYPE PV 1 BIT PV PV PV PV O2_DEV CO2_DEV O2_FAIL CO2_FAIL LOWO2PR LOWCO2PR O2MINLMT O2MAXLMT DOO2ENBL DORPMENBL RPMMINLMT RPMMAXLMT DO_DEV DOOPT_FAIL PHCO2ENBL 9 BEEP_ENBL AUTO_ACK 7 DOCMD 8 AUTOSTART CO2_ON CO2_DEVENBL CO2ENBL O2_ON O2_DEVENBL O2ENBL DOOPTENBL DO_DEVENBL DOOPT_ON

DOCTRL_SW
2

SETDO0%_SW
3

SETDO100_SW
4

SETDOSC_SW
5

SETDOTC_SW
6

CO2RSP_ON
10

O2RSP_ON RPMSP_ON

O2CTRL_SW CO2CTRL_SW

Discrete I/O Table WAVEPOD POD2 (Bay 2&4)

11 12 13

CO2MANUAL O2MANUAL

14 15

71

16

Specifications WAVEPODTM Integrated Instrumentation Unit

Integrated instrumentation unit for dissolved oxygen (optical sensing) and O2 gas mixing controls for use with the System20/50EHT series Wave BioreactorTM. Made of high quality stainless steel and aluminum; compact size and large color touchscreen for ease of use.

WAVEPOD-L version shown

Features:

Color touchscreen Built-in Ethernet interface 4-bay stainless-steel chassis 15.6in x 10in x 14in (397mm x 254mm x 356mm) 19lbs (8.6 Kg) Shipping Weight: 25lbs (11.3 Kg)

Dimensions: Weight: Performance:

RJ12-6 connector (bioreactor base) RJ11 MODBUS jack (Ethernet) DO measurement range 30 100% saturation DO set point range 0 100% saturation O2 set point range 0 50% O2 control range 21 40%, +/- 0.6% 41 50%, +/- 1.0% Environmental:

Airflow set point 0.0 1.0 lpm Airflow control range 0.0 0.5 lpm DO fiber optic probe accuracy +/- 2.0% O2 gas inlet pressure regulated 10 -15 psig (0.7 1.0 bar) O2 gas inlet flow rate minimum of 0.6 lpm

Operating conditions 5 to 40o C. Operating humidity 10 to 90% non-condensing Storage conditions -20oC to +60oC Pollution Degree 1 IP30 rating Indoor Use only 90-240 VAC, 50/60 Hz, 3 A (autoswitching)

Power: Catalog Numbers/ Configurations*:

POD-L0112 DOOPT and O2MIX POD-R0112 DOOPT and O2MIX

L designation = left-side probe connections R designation = right-side probe connections

Options:

Country-specific power cord

WAVEPOD

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WAVEPODTM Integrated Instrumentation Unit

Integrated instrumentation unit for dissolved oxygen (optical sensing), CO2 and O2 gas mixing controls for use with the System20/50EHT series Wave BioreactorTM. Made of high quality stainless steel and aluminum; compact size and large color touchscreen for ease of use.

WAVEPOD-L version shown

Features:

Color touchscreen Built-in Ethernet interface 4-bay stainless-steel chassis 15.6in x 10in x 14in (397mm x 254mm x 356mm) 19lbs (8.6 Kg) Shipping Weight: 25lbs (11.3 Kg)

Dimensions: Weight: Performance:

RJ12-6 connector (bioreactor base) RJ11 MODBUS jack (Ethernet) DO measurement range 30 100% saturation DO set point range 0 100% saturation CO2 set point range 0 15% CO2 control range 0 7.5%, +/-0.38% 7.5% - 15%, +/-0.75% O2 set point range 0 50% O2 control range 21 40%, +/- 0.6% 41 50%, +/- 1.0% Environmental:

Airflow set point 0.0 1.0 lpm Airflow control range 0.0 0.5 lpm DO fiber optic probe accuracy +/- 2.0% CO2 gas inlet pressure regulated 10 15 psig (0.7 1.0 bar) CO2 gas inlet flow rate minimum of 0.2 lpm O2 gas inlet pressure regulated 10 -15 psig (0.7 1.0 bar) O2 gas inlet flow rate minimum of 0.6 lpm

Operating conditions 5 to 40o C. Operating humidity 10 to 90% non-condensing Storage conditions -20oC to +60oC Pollution Degree 1 IP30 rating Indoor Use only 90-240 VAC, 50/60 Hz, 3 A (autoswitching)

Power: Catalog Numbers/ Configurations*:

POD-L0113 DOOPT, CO2MIX and O2MIX POD-R0113 DOOPT, CO2MIX and O2MIX
L designation = left-side probe connections R designation = right-side probe connections

Options:

Country-specific power cord

WAVEPOD

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WAVEPODTM Integrated Instrumentation Unit

Integrated instrumentation unit for pH and CO2 gas mixing controls for use with the System20/50EHT series Wave BioreactorTM. Made of high quality stainless steel and aluminum; compact size and large color touchscreen for ease of use.

WAVEPOD-L version shown

Features:

Color touchscreen Built-in Ethernet interface 4-bay stainless-steel chassis 15.6in x 10in x 14in (397mm x 254mm x 356mm) 19lbs (8.6 Kg) Shipping Weight: 25lbs (11.3 Kg)

Dimensions: Weight: Performance:

RJ12-6 connector (bioreactor base) RJ11 MODBUS jack (Ethernet) pH control range 5 9 CO2 set point range 0 15% CO2 control range 0 7.5%, +/-0.38% 7.5% - 15%, +/-0.75% Environmental:

Airflow set point 0.0 1.0 lpm Airflow control range 0.0 0.5 lpm pH probe range 5 9; accuracy +/- 0.2 pH unit CO2 gas inlet pressure regulated 10 15 psig (0.7 1.0 bar) CO2 gas inlet flow rate minimum of 0.2 lpm

Operating conditions 5 to 40o C. Operating humidity 10 to 90% non-condensing Storage conditions -20oC to +60oC Pollution Degree 1 IP30 rating Indoor Use only 90-240 VAC, 50/60 Hz, 3 A (autoswitching)

Power: Catalog Numbers/ Configurations*:

POD-L1011 pH and CO2MIX POD-R1011 pH and CO2MIX

L designation = left-side probe connections R designation = right-side probe connections

Options:

Country-specific power cord

WAVEPOD

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WAVEPODTM Integrated Instrumentation Unit

Integrated instrumentation unit for dissolved oxygen (optical sensing), pH and CO2 gas mixing controls for use with the System20/50EHT series Wave BioreactorTM. Made of high quality stainless steel and aluminum; compact size and large color touchscreen for ease of use.

WAVEPOD-L version shown

Features:

Color touchscreen Built-in Ethernet interface 4-bay stainless-steel chassis 15.6in x 10in x 14in (397mm x 254mm x 356mm) 19lbs (8.6 Kg) Shipping Weight: 25lbs (11.3 Kg)

Dimensions: Weight: Performance:

RJ12-6 connector (bioreactor base) RJ11 MODBUS jack (Ethernet) pH control range 5 9 DO measurement range 30 100% saturation DO set point range 0 100% saturation CO2 set point range 0 15% CO2 control range 0 7.5%, +/-0.38% 7.5% - 15%, +/-0.75% Environmental:

Airflow set point 0.0 1.0 lpm Airflow control range 0.0 0.5 lpm pH probe range 5 9; accuracy +/- 0.2 pH unit DO fiber optic probe accuracy +/- 2.0% CO2 gas inlet pressure regulated 10 15 psig (0.7 1.0 bar) CO2 gas inlet flow rate minimum of 0.2 lpm

Operating conditions 5 to 40o C. Operating humidity 10 to 90% non-condensing Storage conditions -20oC to +60oC Pollution Degree 1 IP30 rating Indoor Use only 90-240 VAC, 50/60 Hz, 3 A (autoswitching)

Power: Catalog Numbers/ Configurations*:

POD-L1111 pH, DOOPT and CO2MIX POD-R1111 pH, DOOPT and CO2MIX
L designation = left-side probe connections R designation = right-side probe connections

Options:

Country-specific power cord

WAVEPOD

75

WAVEPODTM Integrated Instrumentation Unit

Integrated instrumentation unit for pH, dissolved oxygen (optical sensing), CO22 and O2 gas mixing controls for use with the System20/50EHT series Wave BioreactorTM. Made of high quality stainless steel and aluminum; compact size and large color touchscreen for ease of use.

WAVEPOD-L version shown

Features:

Color touchscreen Built-in Ethernet interface 4-bay stainless-steel chassis 15.6in x 10in x 14in (397mm x 254mm x 356mm) 19lbs (8.6 Kg) Shipping Weight: 25lbs (11.3 Kg)

Dimensions: Weight: Performance:

RJ12-6 connector (bioreactor base) RJ11 MODBUS jack (Ethernet) pH control range 5 9 DO measurement range 30 100% saturation DO set point range 0 100% saturation CO2 set point range 0 15% CO2 control range 0 7.5%, +/-0.38% 7.5% - 15%, +/-0.75% O2 set point range 0 50% O2 control range 21 40%, +/- 0.6% 41 50%, +/- 1.0% Environmental:

Airflow set point 0.0 1.0 lpm Airflow control range 0.0 0.5 lpm pH probe range 5 9; accuracy +/- 0.2 pH unit DO fiber optic probe accuracy +/- 2.0% CO2 gas inlet pressure regulated 10 15 psig (0.7 1.0 bar) CO2 gas inlet flow rate minimum of 0.2 lpm O2 gas inlet pressure regulated 10 -15 psig (0.7 1.0 bar) O2 gas inlet flow rate minimum of 0.6 lpm

Operating conditions 5 to 40o C. Operating humidity 10 to 90% non-condensing Storage conditions -20oC to +60oC Pollution Degree 1 IP30 rating Indoor Use only 90-240 VAC, 50/60 Hz, 3 A (autoswitching)

Power: Catalog Numbers/ Configurations:

POD-L1113 pH, DOOPT, CO2MIX and O2MIX POD-R1113 pH, DOOPT, CO2MIX and O2MIX
L designation = left-side probe connections R designation = right-side probe connections

Options:

Country-specific power cord

WAVEPOD

76

Technical Reference

Optical Dissolved Oxygen

The DOOPT dissolved oxygen (DO) module is used to monitor dissolved oxygen levels in the Cellbag. It can also be used for other bioreactors devices such as spinners. The DOOPT module uses the DOOPT-PROBE optical oxygen measurement probe. The principle of operation is based on the quenching of luminescence of dye molecules immobilized in the probe by molecular oxygen. This is a well known phenomenon, however most instruments that employ this principle determine the amount of oxygen by measuring the intensity of the luminescence signal. This has many disadvantages: 1) the reading is affected by ambient light, 2) the signal diminishes with time due to photobleaching and 3) accuracy is poor in the high oxygen concentration region (> 60% oxygen saturation). These intensity-based devices are low cost but have not proven to be reliable and accurate enough for process monitoring and control of bioreactors. The DOOPT module overcomes these limitations by measuring the phase angle shift caused by the quenching reaction when excited by sinusoidally modulated light. The major advantages of phase-angle based oxygen sensing over conventional Clark-type electrodes are: 1. No oxygen is consumed in the measurement 2. Fast response time (t90 < 10 s) 3. Can measure oxygen in gas or dissolved in liquids 4. Miniature size due to fiber optic technology 5. Long life with little drift 6. Not prone to drying out The DOOPT module provides amplification, display and data transmission of the dissolved oxygen concentration. It allows the real-time measurement of dissolved oxygen concentration inside the Cellbag bioreactors. A special fiber optic DO microsensor is used inside a sealed silicone Oxywell2 sleeve. This allows the probe to be removed for readjustment without compromising sterility. Disposable DOOPT-PROBEs may be reused for many months until photobleaching causes low signal amplitude and the probe becomes progressively too noisy for accurate measurements.

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In the Wave Bioreactor application, the DOOPT-PROBE is used in the proven Oxywell configuration. The Oxywell2 is a silicone rubber sleeve that is provided inside the Cellbag cultivation bag. The DOOPT-PROBE is inserted into the sleeve and oxygen is measured non-invasively as it diffuses through the silicone rubber. The use of the Oxywell increases the response time (t90) from 6 seconds (bare probe) to 3-5 minutes (wet Oxywell). This is still fast enough for the monitoring and control of cell culture. It provides the tremendous advantage of being able to remove, readjust and reuse the DOOPT-PROBE. The silicone sleeve is made of FDA approved Class VI plastic and is the only contact surface. The silicone sleeve eliminates any concern about toxicity of the sensing dye or leaching of the dye into the culture fluid. The bare probe with its fast response time can be used for dynamic oxygen transfer measurements.

DO probe

Oxywell2 sleeve

Cellbag bioreactor
The DOOPT module measures the luminescence decay time of the immobilized luminophore on the tip of the DOOPT-PROBE. The luminophore is typically a transition metal complex of Ruthenium. On excitation by incident light, the luminophore emits light. In the presence of a quencher, such as molecular oxygen, the luminophore becomes deactivated and does not emit light. A relationship exits between the oxygen concentration in the sample and the luminescence intensity as well as the luminescence lifetime. This relationship is described in the Stern-Volmer equation:

= f ([O2 ])
or

o = 1 + K sv [O2 ]
Where o is the luminescence decay time in the absence of oxygen is the luminescence decay time in the presence of oxygen Ksv is the Stern-Volmer constant and depends on the immobilized dye [O2] is the oxygen concentration

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While it is possible to estimate the oxygen concentration by measuring the luminescence intensity, the results are not very reliable. The intensity varies with time as the sensor photobleaches. It is also sensitive to ambient light. In addition, the slope of the intensity response becomes quite shallow with oxygen concentrations greater the 60% and this leads to very poor accuracy and reproducibility above this oxygen concentration. For these reasons, the DOOPT module does not use luminescence intensity in order to estimate oxygen concentration. Instead, the luminescence decay time is measured using a phase-modulation technique. The luminophore is excited with sinusoidally modulated light. The decay time of the luminophore causes a delay in the emitted light signal. This delay is manifested in a shift in the phase angle between the excited and the emitted light signal. The amount of this phase shift is a function of the oxygen concentration as shown in the following equation:

tan 2f mod

or tan = 2f mod

where is the phase angle and fmod the modulation frequency Measurement of the phase shift has several advantages: Variations in the indicator dye luminescence or in the intensity of the light source do not effect the measurement. The decay time is not influenced by bending of the fiber or tip geometry. The measurement is not significantly influenced by the concentration of the dye. This reduces the impact of photobleaching and leaching of the dye. The decay time is not influenced by variations in the optical properties of the sample including turbidity, refractive index and color.

The oxygen measurement is reported at % of air saturation. Air is referenced to 20.95% oxygen. This corresponds to 100% saturation. Since the Stern-Volmer equation is highly non-linear, appropriate linearization calculations are performed by the microprocessor inside the DOOPT module.

Effect of Water Vapor

Water vapor influences the oxygen partial pressure in air and for most accurate results the adjustment should be in air-saturated water or water-saturated air.

Effect of Temperature

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Temperature affects the luminescence decay time as well as the intensity. The diffusion of oxygen is greater at higher temperatures and this also causes variations. In addition, solubility of oxygen in water is temperature dependent and can be described by Henrys law. The DOOPT module compensates for sensor temperature variations as well as solubility effects using a fourth degree polynomial. It is still recommended that adjustments should be done at the same temperature as the process measurement, if possible.

Measurement Range
The oxygen measurement range is from 0 to 250% air-saturation. The phase resolution is better that 0.05o. The accuracy of the measurement is: 1- 30% air-saturation 30 - > 100% 100 - > 250% 0.2% 0.5% 2.0%

Temperature range -10 to 80 oC.

Cross-sensitivity
There is no cross sensitivity for CO2. Increasing the salt (chloride) concentration decreases the solubility of oxygen. This is why it is best to recalibrate the 100% point in the actual culture fluid (in Oxywell2) once the system temperature has equilibrated. Physiological saline is used in the Oxywell2 to minimize any salt concentration artifacts.

Response Time
Bare probe
The bare probe has a response time (t90) of about 4-5 seconds. The following graph shows the response going from air saturation to 0% oxygen solution.
BARE DOPT
120

100

80

%sat

60

40

20

0 0 1 2 3 4 seconds 5 6 7 8 9

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The next graph shows the response time going from 0% oxygen to air saturation. The response time (t90) is about the same (4-5 seconds).
BARE DOPT 2
120

100

80

%sat

60

40

20

0 0 1 2 3 4 seconds 5 6 7 8 9

Wet Oxywell2
When inserted into the Oxywell2 sheath, the probe has a longer response time due to diffusion through the silicone rubber membrane. The major advantage of the Oxywell2 is that the probe does not need to be sterile since it does not contact the culture fluid. This allows for readjustment, replacement and reuse of the probe without any risk of contamination. The response time in the Oxywell2 is greatly improved by filling the tip with physiological saline solution. Oxywell2 sheaths are supplied prefilled with saline. The response time (t90) in the filled Oxywell2 is about 2-3 minutes as shown in the following graph:
WET oxywell 0->100
120

100

80

%sat

60

40

20

0 0 1 2 3 4 minutes 5 6 7 8

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Wet Oxywell 100->0

120

100

80

%sat

60

40

20

0 0 1 2 3 4 5 minutes 6 7 8 9 10

Dry Oxywell2

In a dry Oxywell2 the response time is quite long due to the relatively large amount of stagnant air in the tip that must diffuse through the silicone. The response time (t90) can range from 5 to 6 minutes as shown in the graph below:
DRYOW 0->100
120

100

80

%sat

60

40

20

0 0 1 2 3 4 minutes 5 6 7 8 9

Use of a dry Oxywell2 for oxygen monitoring in bioreactors is not recommended.

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Stability
For long term stability, the probe is only illuminated intermittently. The default sampling frequency is every 10 seconds. The sampling frequency can be changed for dynamic measurements to as low as once per second. The drift due to photo-bleaching is < 0.6% air-saturation per 100,000 measurements. At 10 second sampling this corresponds to 10 days. The Oxywell2 device permits readjustment of the probe without risk of contamination. For optimal performance, the probe should be readjusted every 2-4 days.

Sterilization
DOOPT-PROBEs are supplied non-sterile since they are used in the noninvasive Oxywell2 device. They can be supplied in an autoclavable version for special applications. Please contact WAVE for additional information on custom applications.

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SOFTWARE KNOWN ISSUES

Overview Screen Kg display is shown when load cell is not present and the option is turned off. Pressing on the area by the O2MIX PV display changes the CO2MIX PV display into the AIR PV display. Pressing on the area by the original AIR PV display changes the new AIR PV display back into the CO2MIX PV display. Retry message appears occasionally even though the unit is set to not communicate with a Base. Acknowledging alarms on the WAVEPOD does not turn the BASE touch screen from red to green. OUT HI PR ALM appears as BLUE (acknowledged) in alarm log. EHT EMERSTOP does not cause alarm button on WAVEPOD to blink RED all the time. If Base type is set to NONE, but the WAVEPOD is connected to a Base unit, alarms from the Base will still appear in the WAVEPOD alarm log. Black box blinks onto the screen for < 1 sec. Occurs more infrequently than before. An incorrect keypad can be accessed by pressing most keypad buttons. When exiting the password protected area quickly and then re-entering that area again quickly, the system does not prompt user for a password. Latency or Maple issues. System screen resets occasionally. Pressing on the area where the alarm button appears switches the screen to the alarm log even if the alarm button is not present. No action required by user.

Overview Screen

The user can switch to a different screen and return in order to view the proper CO2MIX PV display.

Overview Screen

No action required by user. User should acknowledge alarms on the BASE unit in order to change the touch screen state. No action required by user. No action required by user. User should disconnect the communication cable between the WAVEPOD and the BASE if the Base type is set to NONE. No action required by user. Press ESC to exit the keypad and try accessing it again. User can wait 10 seconds before accessing the password protected area again. Issue may arise when resetting the unit using the RESET button. If the problem occurs frequently, call service for assistance. No action required by user.

Alarms Alarms Alarms

Miscellaneous

Miscellaneous

Miscellaneous

Miscellaneous

Miscellaneous

Miscellaneous

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WAVEPOD

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For contact information for your local office, please visit www.gelifesciences.com/contact GE Healthcare Bio-Sciences AB Bjrkgatan 30 751 84 Uppsala Sweden www.gelifesciences.com/bioprocess GE Healthcare Bioscience BioProcess Corp. 300 Franklin Square Drive Somerset, NJ 08873 USA www.gelifesciences.com/wave

GE, imagination at work and GE monogram are trademarks of General Electric Company. WAVE Bioreactor is a trademark of GE Healthcare Companies. 2008 General Electric Company All rights reserved. First published March 2008. All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information. Wave Europe Pvt. Ltd. Unit 10, Mathew Hill Commercial Park Forge Hill, Kinsale Road Cork, Ireland GE Healthcare Bio-Sciences AB Bjrkgatan 30 751 84 Uppsala Sweden GE Healthcare Europe GmbH Munzinger Strasse 5, D-79111 Freiburg, Germany GE Healthcare UK Ltd Amersham Place, Little Chalfont, Buckinghamshire, HP79NA, UK GE Healthcare Bio-Sciences Corp. 800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ 08855-1327, USA

GE Healthcare Bio-Sciences KK Sanken Bldg, 3-25-1 Hyakunincho, Shinjuku-ku, Tokyo 169-0073, Japan

450032 F

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