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Instruction Manual

8.2.5.3 Adding Base
After inoculation, an alkaline like Sodium Bicarbonate or Sodium Hydroxide, may be automatically added to the MCV to maintain pH. This
procedure shows how to connect the Base Bottle to automatically add the base solution to the MCV.
NOTE:

Both the Base and Feed pumps only run in one direction, counterclockwise, to advance flow to the MCV.

1. Connect Base tubing line to the MCV.

• Locate the base tubing line (yellow and blue clips) on the MCV headplate.

•  Use the AseptiQuik® S connections attached to the Base dip-tube and base addition line.
•  Connect the AseptiQuik® S connectors to an audible click.
•  Follow AseptiQuik® connection procedure in Section 8.2.1.2.

2. Tubing install in base pump.

• Load base tubing in the base pump. Install tubing line according to the colors indicated on the

base pump cover.

• Twist the gray lever to open and close the pump head.
TIP:

Make sure tubing in the pump head is centered between the V-groove clamp.

• This illustration shows the base bottle set up to automatically add base to the MCV.

3. Prime Base tubing in Manual mode.

•  On the HMI, select the base pump icon. Press Program to run in Manual mode.
•  Press the Operator icon.
•  Stop the pump immediately before base enters the MCV (approximately 15 to 20 mL).
•  Return the base pump to program mode.

8.2.6 Media Exchange Phase
The user selects the volume of media from the MCV for removal and replaces it with fresh media. This phase differs from Media Addition
and Media Removal Phases since it allows the user to partially drain or completely drain the bioreactor in addition to the MCV, and
upon restart minimizes transients in pH, DO, and temperature. The user shall ensure that there is sufficient media addition to refill the
bioreactor and maintain the minimum fill level in the MCV.

1.

On the HMI, select Media Phases.
Select Media Exchange.

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Содержание Ascent 6970

Страница 1: ...Corning Ascent Fixed Bed Reactor Process Development System Instruction Manual Catalog Number 6970 ...

Страница 2: ... MCV and FBR Heaters 17 5 5 1 MCV Heater and Temperature Sensor 17 5 5 2 FBR In line Heater and Temperature Sensor 18 5 6 Bar Code Scanner 18 6 0 Process Overview 19 6 1 System Set Up 20 6 1 1 Initialization 21 6 1 2 Media Fill and Prime Phase 21 6 1 3 Media Conditioning Phase 21 6 1 3 1 Sensor Calibration 21 6 2 Cell Culture 22 6 2 1 Media Maintenance 22 6 2 2 Inoculation 22 6 2 3 Media Exchange ...

Страница 3: ...ss 59 7 13 2 Remote Access to Data File Management 59 7 14 Alarms 60 7 14 1 No Alarm Interlocks 62 7 15 Power Loss and Recovery 63 8 0 Protocols 64 8 1 Pump Calibration 66 8 2 Media Phases 68 8 2 1 Initialize Phase 68 8 2 2 Media Fill and Prime Phase 76 8 2 3 Media Conditioning Phase 79 8 2 3 1 Sensor Calibration 80 8 2 4 Media Maintenance Phase 81 8 2 5 Inoculation Phase 82 8 2 5 1 MCV Inoculatio...

Страница 4: ...Enclosure Filter Replacement 114 10 0 Warranty 116 11 0 Equipment Disposal 116 Appendix 1 PID Loop Calculations and 117 Tuning Guidance Appendix 2 Phase Permissive 118 Appendix 3 Operational Sequence Reference 119 Tables 1 Media Phases 119 2 Auxiliary Phases 128 3 Harvest Phases 132 Appendix 4 Quick User s Guide 134 Set Up 134 Experiment Run 135 Harvest Set Up 136 ...

Страница 5: ...ing labels Warning Label Description High voltage symbol To reduce the possibility of injury turn off power and unplug before servicing Biological hazard symbol Adhere to proper PPE when handling media Hot surface symbol Avoid touching the following heated surfaces Media Conditioning Vessel MCV and bioreactor nests Exhaust filter heater Lock out power and allow surface to cool before servicing Hea...

Страница 6: ...s Figure 2 1 and Figure 2 2 During cell culture a recirculation pump circulates media from the MCV through the FBR A separate pump aids in removing depleted media and replacing the MCV with fresh media during media maintenance Cell culture pH and nutrient levels are monitored and maintained via feed and base pumps while temperature controlled heated nests keep the MCV and bioreactor at a constant ...

Страница 7: ... Indoor temperature range is 5 C to 40 C For optimal use the indoor temperature range shall be between 20 C to 30 C Maximum relative humidity at 80 for temperatures up to 30 C Maximum altitude up to 2 000 m Pollution degree 2 Up to IEC Overvoltage Category II for transient over voltages Space Required A workbench suited to accommodate the equipment dimensions L x W x H 53 x 30 x 27 in 134 62 x 76 ...

Страница 8: ...the Corning Ascent FBR instrument 4 1 Power Air and Gas Connections The Corning Ascent FBR is a benchtop instrument with accessible connection ports to main power air O2 CO2 and N2 Figure 4 1 Push the On Off LED button along the front left base of the instrument Figure 4 2 to start up the HMI The LED button will illuminate green when power is on Allow the system to warm up for 30 minutes before st...

Страница 9: ...feed and base bottles 5 1 1 Media Addition and Media Waste Bottles Fresh cell culture media used during media fill and exchange is filled in the media addition bottle on the side of the Ascent instrument Figure 2 2 Media is channeled to the Media Conditioning Vessel MCV via tubing and the media pump More than one media addition bottle can be connected via AseptiQuik connectors as needed During med...

Страница 10: ...n draining the bioreactor e g at substrate sampling and media exchange Valve default position is closed T5 Channels transfection agent and inoculum aseptically to the MCV T6 Tubing line with red and blue clips to the feed bottle Feed is added to the MCV via the feed pump T7 Tubing line with yellow and blue clips to the base bottle Base is added to the MCV via the base pump p1 Auxiliary Luer Lock p...

Страница 11: ...rculation pump tubing The two pieces of media tubing that connect the MCV to the bioreactor via sterile connectors are clamped across the tray along with two in line DO sensors and an in line MCV pH sensor The sensor tray serves the following purposes Allows easy load and installation of the pump tube in the recirculation pump Allows easy color identification to connect the sensors that monitor MC...

Страница 12: ...e MCV 5 1 5 FBR with Sterile Connectors The Corning Ascent FBR system has two options to connect media flow from the MCV to the FBR and return media from the FBR to MCV Option A uses Lynx connectors while Option B uses AseptiQuik G connectors Figure 5 6 and Figure 5 7 At experiment start the sterile connectors Option A or B are connected to their mating sterile connectors on the MCV The Lynx conne...

Страница 13: ... be removed each day for sampling 5 1 6 Harvest Kit The Corning Ascent FBR system has a consumable harvest kit in a portable tray for easy positioning enabling users to recover cells from the bioreactor Figure 5 10 Prior to harvest the bioreactor is fluidically isolated from the MCV by clamping the recirculation inlet and the outlet tubing During the Harvest Phase the recirculation pump in conjunc...

Страница 14: ...or before introducing harvest solution This bottle is also used after introducing harvest solution at post harvest wash Some components of cell culture media may inactivate the harvest solution active enzymes Hence it is recommended to perform a bioreactor DPBS wash before introducing harvesting solution F Flush The bottle holds a predefined volume of cell dissociation reagent Accutase used to flu...

Страница 15: ...le 1 Waste media collection MCV to media waste bottle 2 Waste media collection Base bottle to MCV Base addition Feed bottle to MCV Feed addition Alternate connection to inoculum bottle Alternate site for seeding the bioreactor Tubing line on MCV Inoculum addition Tubing line on MCV Transfection addition to MCV Table 5 6 Sterile connections at Harvest set up AseptiQuik G AQG17004 AseptiQuik DC AQCD...

Страница 16: ...he MCV gas vent Section 5 2 3 4 MCV sparge Sparging gas line in the MCV Section 5 2 4 5 MCV temperature In line sensor monitors the media temperature exiting the MCV via the recirculation loop Section 5 2 5 6 FBR DO DO2 Monitors dissolved oxygen levels exiting the bioreactor Section 5 2 6 7 MCV pH Monitors pH levels exiting the MCV Section 5 2 7 8 MCV DO DO1 Monitors dissolved oxygen levels exitin...

Страница 17: ...egulated and controlled in the software settings for Media Conditioning The gasses flow through a sterile filter in the MCV The gas mixture composition O2 CO2 air N2 in the cell culture media is automatically controlled by the system to maintain user defined oxygenation and pH levels The gas sparge line has a positive shut off valve that closes to ensure no leakage when the mass flow controller MF...

Страница 18: ...is automatically maintained via the pH controller which controls two different outputs the base pump flow rate and the CO2 flow rate SCCM in sparging gas mixture pH sensor calibration values are provided by the manufacturer and can be automatically read into the system by bar code scanning the MCV Consumable set or manually entered by the user Figure 5 21 MCV pH sensor monitors media pH level 5 3 ...

Страница 19: ...se bottle to MCV Pump speed approximately 0 1 to 80 SCCM calibration dependent Adds base to the MCV for pH maintenance Table 5 8 Peristaltic pump descriptions Figure 5 23 The recirculation pump uses robust tubing to withstand continuous pumping Figure 5 24 Base and feed pumps have color coded covers to match the colored clips on the tubing lines 5 4 Pinch Valves There are five pinch valves Figure ...

Страница 20: ...osition denoted by a black bar on the pinch valve cover is farthest from the instrument The bottom valve position denoted by a white bar on the pinch valve cover is closest to the instrument When the system powers on the pinch valves are not energized At the de energized state the bottom valve plunger is designed to extend while the top valve plunger is retracted Figure 5 27 Tubing load takes plac...

Страница 21: ...ctor to harvest collection bottle Channels harvest flush to the harvest solution bottle during harvest solution recirculation Harvest Solution bottle Harvest Collection bottle The blue pinch valve channels flow from the bioreactor to the harvest waste bottle or to the harvest flush bottle Harvest Waste bottle Harvest Flush bottle 5 5 MCV and FBR Heaters This section describes the heated MCV and FB...

Страница 22: ...range between 30 C to 40 C The bioreactor heater is limited to 80 of the duty cycle to prevent overheating Figure 5 29 Cross section of the bioreactor in the tray and the Lynx connectors in the insulated heated nest Figure 5 31 Illustration showing the temperature sensor plugged in while the bioreactor is in use Figure 5 32 Bioreactor temperature sensor location in the media tubing along the tray ...

Страница 23: ...ds There are two types of phases Sequence Phase Media and Harvest and Auxiliary Phase Some phases can only run once while other phases can run multiple times Media Phases can run after a Harvest Preparation Phase but not during an active Harvest Phase Auxiliary Phases can be initiated during an active Media Phase or an active Auxiliary phase and will put the active phase on hold see section 6 3 Ta...

Страница 24: ...moval Final Rinse Once Harvest Preparation is initiated these Harvest Phases can run multiple times to detach cells from the substrate and collect the cells in the harvest collection bottle Auxiliary Phases Auxiliary Phases cannot run while Harvest Phases are active 6 1 System Set Up At system set up the user starts the Initialize Phase The consumables bar codes are first scanned and then the vess...

Страница 25: ...lation pump This phase maintains the Media Phase setpoints shown in Table 6 3 Temperatures are automatically controlled to user setpoints The Stabilization duration allows sufficient time to reach steady state prior to calibrating sensors typically 2 to 4 hours or more Table 6 3 Media conditioning setpoints Setpoints Agitator Recirculation Pump MCV Temperature FBR Temperature Air MFC Flow Rate O2 ...

Страница 26: ...et up over the ramp time specified The system is now at a steady state for cell expansion and the recirculation pump flow rate will be automatically adjusted to maintain the FBR DO reading to its setpoint or if the actual FBR DO is above the setpoint the minimum pump speed will be maintained Table 6 6 shows the setpoints controlled Table 6 6 Inoculation setpoints Setpoints Recirculation Attachment...

Страница 27: ...ing Air Flow Rate is set to maintain DO at a favorable level The bioreactor is then manually removed to a biosafety cabinet for substrate sampling Table 6 9 shows the setpoints controlled After the bioreactor is reinstalled the system auto starts via Stabilization Time to the previous setpoint to minimize the control loop transients in the system The Pump Ramp Duration controls the time required t...

Страница 28: ...e 6 13 shows the setpoints controlled Table 6 13 Perfusion or media replacement setpoints Setpoints Media Pump Addition Flow Media Pump Removal Flow Media Volume Number of Cycles Perfusion Cycle OFF Duration 6 3 6 Media Dilution This phase continuously recycles media by transferring media to the media bottle from the MCV and back to the MCV from the same media bottle for a specified number of cycl...

Страница 29: ...gure 6 4 shows a simplified view of the harvest wash path Table 6 15 shows the setpoints controlled Table 6 15 Harvest wash setpoints Setpoints Harvest Wash Recirculation Pump Flow Rate Harvest Wash Volume 6 4 3 Cell Release Accutase or a similar proteolytic cell dissociating enzyme is flown through the bioreactor for cell release Solution from the harvest solution bottle is pumped through the bio...

Страница 30: ...collection bottle during cell removal When pressure is applied to the flush bottle for the Removal Duration harvest solution is flowed at around 2L min through the bioreactor into the collection bottle The goal of fast flow through the bioreactor is to retrieve most of the cells into the collection bottle Figure 6 6 highlights the cell removal path Table 6 17 shows the setpoints controlled NOTE Ce...

Страница 31: ...Table 6 18 Final rinse setpoints Setpoints Final Rinse Recirculation Pump Flow Rate Final Rinse Recirculation Pump Totalizer Volume Figure 6 7 Final rinse path highlighted 7 0 Human Machine Interface HMI This section describes the HMI features and functionalities 7 1 Main Screen Overview Figure 7 1 shows the Cell Culture Process screen which is also the main screen at startup Table 7 1 lists the n...

Страница 32: ...iagnostics Section 7 12 9 VPN Enabled or Disabled VPN has a configurable time limit up to 24 hours after which VPN is automatically disabled Section 7 13 10 These are buttons to navigate and reset alarms Section 7 14 11 Phase status display information with green flashing prompt button an Abort button and Global Reset button Section 7 5 12 System screen for setting up the time and entering the ins...

Страница 33: ...Bypass is enabled and a value is simulated This option is only available at the Admin level Upside down triangle with wrench Bypass is enabled This option is only available at the Admin level Two stacked squares and a rectangle on the side Status indicator Value is held at the last good value Device status symbols Shaded gray Device state color shows that the component is Off disabled Colored whit...

Страница 34: ...ondition has been cleared but is not acknowledged Acknowledgement is required Black alarm bell Alarm condition is active for any of the following MCV DO FBR DO MCV pH MCV Temp and or FBR Temp Flashing red border with double exclamation marks in a red circle Alarm limit indicator Input failure Single down arrow inside a black box Low deviation threshold has exceeded for any of the following MFCs PC...

Страница 35: ...eens Sys button for software information and clean screen Observe historical trends Sys button for user interface shutdown Initiate and Cancel Phases Global Abort button use Change probe calibration Pump calibration Pressure zero point adjustment during Initialize Phase Data button Data File Entry Meta data Entry Recirc Volume Change Date and Time View Alarm History Password Admin button to reset ...

Страница 36: ...l a full unit power cycle Figure 7 6 Shut down user interface 7 4 System Screen Selecting the Sys button next to the date and time field will show the software information screen Figure 7 4 The system ID name in the field is displayed on each HMI screen There are four options on the Sys window Clean Screen Time and Date Shutdown User interface and Password Admin Time and Date and Password Admin ar...

Страница 37: ... Reset Operator Password 1 While logged on as Engineer select the Sys button and then the Password Admin button 2 Select the Login Operator button enter the Operator login information and then accept 3 Select the Reset Operator Password button 4 Enter current password and new password then accept 5 Login Engineer can then be selected and login information can be entered to return to Engineer the a...

Страница 38: ... 0 except for the pH equipment module which is set to 50 The vent filter heater remains on All pinch valves except for the pressure control valve PCV and the harvest pressure valve de energize after 3 seconds to prevent pressure build up from pumps ramping down The pressure control valve and the harvest pressure valve de energize immediately Figure 7 9 Abort and Global Reset buttons After selectin...

Страница 39: ...ance Puts DO pH and temperature loops into Automatic mode at user setpoints Once triggered Media Maintenance Phase cannot be cancelled Inoculation Starts inoculation timer prompts for volume recirculates at attachment speed and time Media Exchange Replaces media in system per set up Transfection Mixes for a set time updates MCV volume and restarts loops Shutdown Stops all loops and stops data logg...

Страница 40: ...val flow rate independent of addition flow rate and delay between systems to minimize transients Table 7 7 Harvest Phases descriptions Harvest Phases Description Harvest Preparation All loops heaters DO and pH are turned off Preparation time to connect Harvest kit to the bioreactor and fluidically isolate the bioreactor Harvest Preparation can only be performed once Harvest Wash The bioreactor is ...

Страница 41: ...the Media Phase will immediately be put on hold and will resume once all active and held auxiliary phases are completed Should the user trigger a Media Phase while another Media Phase is active the active Media Phase will be canceled and the new Media Phase will begin immediately The faceplate removes the play symbol after a phase is completed Figure 7 13 The phase button on the HMI is grayed out ...

Страница 42: ...int Likewise in line sensor probes can be calibrated via Probe Calibration Figure 7 15 DO and pH probe values are bar coded into the system The user either verifies the bar coded values or manually enters values from the data sheet during Media Conditioning see Section 8 2 3 1 Temperature offsets can be determined offline in manual operation on a non sterile system Defaults were determined in a 22...

Страница 43: ...aph 8 Controls 4 Alarm see Table 7 20 9 Edit button and opens the Deviation Alarm Editor 5 Reset 7 8 1 Equipment Parameter Control Faceplates Selecting an equipment component e g pinch valve will create a pop up with the corresponding tag name current state setpoint actual value and command options Figure 7 17 shows all equipment faceplates that allow control access options listed in Table 7 9 Fig...

Страница 44: ...oop modes change loop setpoints when in Operator mode and to configure and tune the control loop Each of the main control loops pH DOs temperatures has one of these faceplates NOTE All PID loops can be run in Operator Auto and Manual modes from the PID faceplate The temperature loops are configured from the faceplates while the two DO loops and the pH loop are configured from the Configuration But...

Страница 45: ... Configuration window for alarms and loop tuning Figure 7 22 Graphical tuning window Figure 7 23 Graphing tuning window and PID coefficient window Figure 7 22 is the PID tuning graphic used to monitor loop parameters during PID tuning Figure 7 23 shows the PID parameters for control These are preconfigured and should only be adjusted by a PID loop tuning expert Use Appendix 1 for loop calculation ...

Страница 46: ... each output actual values at the current time 4 The P I and D gains control how quickly the loop acts to adjust the DO CV value if the measured DO deviates from the user setpoint NOTE The vast majority of applications will be best served by leaving the D Gain at zero for the MCV DO controller configuration NOTE If user inputs Total Flow setpoints that exceed the systems ability to deliver the des...

Страница 47: ...Control Loop Configuration example values when gas flow is held constant Figure 7 27 MCV DO Control Loop Configuration example values when each parameter is cascaded stepwise Figure 7 29 MCV DO Control Loop Configuration example values when flow and agitator speed rpm are constant and O2 is non linear piece wise linear between Controller Value points ...

Страница 48: ... to base flow rate The Split Range Control graph in Figure 7 32A illustrates how the control loop works PID CV is 50 when the loop is at its minimum output This is within the default split range Controller Value scaling region wherein both the CO2 MFC flow rate and base pump flow rate are set to their minimum values By default the split range Controller Value scaling limits are 49 9 and 53 Base Pu...

Страница 49: ...O2 MFC flow rate is set to the appropriate value in accordance with the minimum CO2 setpoint entered in the Media Maintenance Phase Region B shows when the pH loop is above the upper deadband limit The loop is in automatic control and the Controller Value is limited to a maximum of 50 Region C shows when the pH is below both the setpoint and the lower deadband limit The loop is in automatic contro...

Страница 50: ...rly noisy causing undesirable jumps in the pump speed if the D Gain is non zero Figure 7 33 Bioreactor DO Control Figure 7 34 Bioreactor DO Loop Configuration Control loop output Controller Value 7 10 Accessing Trend Properties The trend page Figure 7 35 displays trends and behavior of different parameters over a selected period up to the full batch processing time The four trends Main Process DO ...

Страница 51: ...hart area to change the zoom After the box is drawn the chart is resized to show only the selected area 8 Statistics Observe statistics for all traces on the page Minimum Maximum Average Std Dev Allows to print table 20 Alarm Cursor Not in use 9 and 10 Display previous or next trace on the Y axis if not stacking 21 Stack Event Y axes Not in use 22 Save Template Saves the current template If change...

Страница 52: ...tton to refresh the Chart To set a relative time period Click the arrow next to the third drop down box Then select the relative time period 7 10 2 Tag List Features The tag list displays all the current tags or traces that are in the current template The tag list is normally hidden To expand the tag frame click on the Tags box Figure 7 38 Figure 7 37 Grayed out versus not grayed out time bar Figu...

Страница 53: ... of traces Style Displays the color and style of the trace line for a given tag Axis Min Minimum Y axis value for the tag Axis Max Maximum Y axis value for the tag Unit Gives the units for the tag Precision Decimal precision for the tag value Click the down arrow to change the number of decimal places displayed Format Indicates if scientific or decimal number format is being used User may click th...

Страница 54: ...laced the values for all displayed tags are shown next to the cursor Figure 7 43 Likewise when the Y axes cursor button is selected two horizontal lines are generated on the chart area that can be moved along the Y axis Figure 7 44 Each cursor can be moved individually Where the Y axis cursor is placed the values for all displayed tags are shown next to the cursor ...

Страница 55: ...ow Rate PV Base Pump Speed PV Base Pump Total Run Volume CO2 in Sparging Gas Mixture 4 Temperature Control FBR Temperature setpoint CV and PV MCV Temperature setpoint CV and PV Recirculation Pump Speed PV 5 Main Process MCV pH PV MCV Temperature PV FBR Temperature PV Agitator Speed PV Recirculation Pump Speed PV FBR DO PV MCV DO CV and PV Figure 7 45 Apply Template dialog box 7 10 4 1 How to Creat...

Страница 56: ...e or enter the same name as the template file they wish to overwrite 3 Press Save in the dialog box to keep the changes made to the existing template 7 10 4 4 Add Tags to Template 1 Click on Automatic in the top right hand corner of the chart area to bring up the Chart Properties dialog box Figure 7 46 Figure 7 46 Press Automatic to pull up Chart Properties box Figure 7 47 Select Traces tab 2 Sele...

Страница 57: ... the green plus button 3 Select the green plus button in the lower left hand corner of the dialog box Figure 7 48 4 Select the small plus button next to Data Logs then select FTDatalog Figure 7 49 Figure 7 49 Select small button and FTDatalog ...

Страница 58: ... now removed Close dialog box to return to the chart area 7 10 4 6 Zoom and Navigate Data Zoom in on a Trend Double click an area of the plot surface Press CTRL plus on the keyboard At the bottom right of the plot surface click the plus sign button Scroll the mouse wheel up This will zoom in relative to the cursor position Zoom out of a Trend Do Shift click an area of the plot surface you want to ...

Страница 59: ...d more Chart User may set up trend chart title information displayed in tool tips zoom in and out and other properties of the trend Chart area X axis User may set up trend horizontal axis Application User may set up trend line styles and trace dimming Retrieval User may set up the retrieval mode and retrieval information displayed on the trend Traces The user may set up the appearance and other op...

Страница 60: ...at the beginning of the run These entries could correspond to the names in the Content Notes fields on the left The metadata file is saved as a CSV file Figure 7 54 and named in the following format DDMMMYYYY_BatchData_Run Name csv The Raw Barcode contains the data as read by the bar code reader or entered by the user manually The Barcode Name represents the name generated by the bar code reader f...

Страница 61: ...box during bar code scanning 7 11 4 USB Save The USB button provides users a way to transfer batch and process data log files from the Ascent instrument to an external data storage See Figure 7 58 and Table 7 18 Table 7 18 Batch and process data files transfer to a USB drive Label Description 1 Stored data path 2 Each batch process generates two files Batch Data File Batch Data file name structure...

Страница 62: ...ton is green The user specifies a time limit for VPN access in the pop up box The button remains green until the time limit is reached When VPN is disabled access to the GUI and data files is still available to the end user from behind the firewall VPN enabled only grants access to the GUI and for software support from outside the firewall Figure 7 60 VPN Enabled button and pop up box 7 12 Diagnos...

Страница 63: ...llation Password length is up to 7 alphanumeric characters Figure 7 62 Figure 7 62 TightVNC remote access and password screens 7 13 2 Remote Access to Data File Management To perform data file management select the data transfer icon in the top ribbon of the TightVNC application Figure 7 63 and follow the steps below Figure 7 64 1 Select the DataStorePlus folder 2 Sort data files by Modified Data ...

Страница 64: ... control faceplate Pressing the bell symbol Figure 7 65 at the bottom of the main screen opens the alarm history which is accessible at Operator and Engineer levels Alarms are recorded on the system with a generation timestamp User acknowledgment is recorded Table 7 20 is a general guide on how to clear alarms Figure 7 64 TightVNC file transfer steps Figure 7 65 Alarms banner and navigation button...

Страница 65: ...int This alarm is not active during the Media Exchange N A User receives warning and is prompted to initiate Media Addition Current MCV Volume falls below LOLO Alarm setpoint This alarm is not active during the Media Exchange Phase Current phase moves to Hold User is prompted to initiate Media Addition User must press Confirm to continue the current phase or initiate Abort Device Faults HIHI Tempe...

Страница 66: ...as inlet valve The gas shutoff valve energizes opens when any of the mass flow controllers are active or at non zero setpoint and closes when all mass flow controllers are at zero setpoint Automated DO pH and temperature control When the recirculation pump is not running automated DO pH and temperature control are not active Automated MCV temperature control When the agitator motor is not running ...

Страница 67: ... FBR temperature HIHI alarm is active Pressure HIHI alarm is active MCV DO EM MCV DO sensor failed Any MCV DO sensor fault active Air MFC fault CO2 MFC fault Gas shutoff valve fault Agitator fault Pressure HIHI alarm is active FBR DO EM FBR DO sensor failed Any FBR DO sensor fault active Recirculation pump is interlocked Pressure HIHI alarm is active pH EM Any pH sensor fault active MCV media HIHI...

Страница 68: ...2 5 Media Exchange Section 8 2 6 Transfection Section 8 2 7 Shutdown Section 8 6 Harvest Harvest Preparation Section 8 4 1 Harvest Wash Section 8 4 2 Cell Release Section 8 4 3 Cell Removal Section 8 4 4 Final Rinse Section 8 4 5 Shutdown Section 8 6 Auxiliary Substrate Sampling Section 8 3 1 Media Addition Section 8 3 3 Bolus Addition Section 8 3 4 Media Removal Section 8 3 2 Perfusion Section 8 ...

Страница 69: ...y start automatically Figure 8 2 Press the flashing green status box to view outstanding tasks Figure 8 3 Secure AseptiQuik connection to an audible click Figure 8 4 Pull out both tabs after AseptiQuik connection is made Below are tips for running a seamless experiment Tip 1 Make sure the correct bioreactor size is selected Tip 2 Tightly mate AseptiQuik connections to an audible click Figure 8 3 T...

Страница 70: ... 700 100 mL min Recirculation LS17 PharmaPure tubing 700 100 mL min Base LS16 platinum cured silicone tubing 90 100 mL min Feed LS16 platinum cured silicone tubing 90 100 mL min Pump Calibration 1 Install the correct pump tubing for calibration see Table 8 3 NOTE Make sure the pump tubing is correctly centered between the pump head V groove clamp 2 Fill an appropriate container with DI water for c...

Страница 71: ...to fill the line with calibration solution Jog until all air has been removed from the calibration tubing and replaced with DI water Tip To activate a pump jog tap and hold the jog button 6 Place the calibration tubing outlet end into an appropriately sized graduated cylinder or bottle This example shows a calibration tubing loaded in the media pump 7 Enter Pump Speed and Seq Duration 1 Select Sam...

Страница 72: ...should be performed to verify pump calibration is valid e g run for 1 minute at a specified flow rate with the pump in Operator mode 8 2 Media Phases This section shows the manual tasks and the process setpoints to enter for each Media Phase 8 2 1 Initialize Phase At Initialization the user enters experiment details confirms that mass flow controllers and pumps are turned off tares the pressure se...

Страница 73: ... MCV in nest Align the MCV base so that it sits snugly in the nest 7 Orange pinch valve set up Locate the orange pinch valve to the right of the media pump Locate the media tubing lines with orange white and orange back clips Load tubing with the white clip into the bottom valve position next to the instrument then load tubing with the black clip in the top valve position farthest away from the in...

Страница 74: ...lter heater Install the exhaust filter in its wire support so that it is upright Press the buttons to secure the heater blanket around the filter 12 Close the valve on the media return line Check that the valve on the media return line is closed Turn twist the media return line to properly orient the tubing lines 13 Untangle the Feed and Base lines Cut zip ties on the Feed and Base tubing lines to...

Страница 75: ...e to seat the fiber optic guide will result in unreliable measurements Close the clamp at the AseptiQuik G connection above the sensor tray Make sure the clamps at the sensor tray are not causing kinks in the media lines It may be necessary to turn twist the sensor tray if the media lines are crossed The sensor tray and interface panel connections are now set up Proceed to connect the MCV and FBR ...

Страница 76: ...holder and is not tilted Check for kinks in the tubing lines Straighten out twists and kinks The consumables are now set up Return to the HMI for further instruction 21 MFC and pumps turned off A prompt will remind the user to complete setup and to confirm that mass flow controllers and pumps are turned off Press to confirm 22 Zero Pressure Sensor Use the numbers on the images to perform the follo...

Страница 77: ...n begins the Elapsed Time counter begins signaling batch start Upon completion of Initialization step Batch start timer starts Proceed to Media Fill Prime phase described in Section 8 2 2 8 2 1 1 Lynx Connections This section shows how to connect sterile lines between the MCV and FBR for instruments using Lynx connectors A video can also be found on https youtu be _QXy37zCES4 1 Locate MCV lines wi...

Страница 78: ...il the top clip is engaged 6 Slide the blue door away from the sterile housing to a locked position 7 Push and twist the Lynx connector blue handle towards the sterile housing TIP To prevent the connected lines from being twisted after connection pre twist in the opposite direction 8 Keep twisting and pushing the blue handle until the notches along the blue handle sit flush with the ones on the st...

Страница 79: ...n to Section 8 2 1 8 2 1 2 AseptiQuik G Connections This section shows how to connect tubing lines between the MCV and FBR for instruments using AseptiQuik G connectors These steps also apply when connecting other AseptiQuik lines e g media addition base and feed connections A video demonstration can be found here https www youtube com watch v XEFy0cQ6cJg 1 Locate the MCV lines with AseptiQuik G c...

Страница 80: ...actor nest Make sure to loop the tubing under the tray ensuring that there are no kinks in the lines especially in the segment from the T connector to the FBR media inlet line The sterile connections between the MCV and FBR are now set up Continue setting up the FBR return to Section 8 2 1 8 2 2 Media Fill and Prime Phase This section shows how to fill the MCV and begin bioreactor priming 1 Fill t...

Страница 81: ...ia addition bottle is aseptically connected and ready for batch and prime On the HMI select Media Phases Select Media Fill Prime 3 Media fill Enter setpoints Media Flow Setpoint Media Fill Volume Recirculation Pump Flow Setpoint Press to start Media Fill NOTE The FBR size is automatically determined from the FBR bar code reading and will be stored in the BRX Loop Volume under the Data button Media...

Страница 82: ...he prompt The recirculation pump automatically switches to Operator mode white flag next to the pump so that the user can begin FBR de airing Follow steps for de airing the FBR 1 Press Pump icon 2 Verify that the mode button says Operator 3 Enter Setpoint 4 Jog forward and backward several times and verify that bubbles are cleared from system TIP Run the recirculation pump in Operator mode for app...

Страница 83: ...edia flow from the MCV to the FBR and returning to the MCV 10 Media prime updates The MCV level will update after priming System Volume MCV Volume BRX Loop Volume 11 Press to confirm Media Fill Prime phase is completed 8 2 3 Media Conditioning Phase This procedure shows how to stabilize temperature DO and pH for Media Conditioning During this phase probe calibrations are performed It is recommende...

Страница 84: ...tioning approximately 2 hours minimum after wetting the in line sensors when the system is stable This procedure shows how to calibrate the in line sensors DO pH and temperature sensors 1 On the Probe Calibration pop up are 1 Bar coded from the MCV consumable kit or manually entered DO and pH calibration parameters Both DO sensors will use the same calibration factors entered 2 Probe calibration r...

Страница 85: ...oratory i e room temperature and 20 70 RH Enter an offset from nominal to compensate for ambient effects MCV temperature offset is typically 2 C to 4 C BRX Temperature Offset is typically 0 C to 1 C Acknowledge the prompt once sensor calibration is complete Press to confirm Media Conditioning completion 8 2 4 Media Maintenance Phase This procedure shows how to maintain proper media conditions in t...

Страница 86: ...modules in Automatic mode according to setpoints Once the system is stabilized after media conditioning proceed with Inoculation described in Section 8 2 5 8 2 5 Inoculation Phase This procedure shows how to add inoculum after the bioreactor is primed 1 Scan the inoculum consumable to enter the bar code 2 Prepare inoculum bottle Fill the inoculum bottle in the biosafety cabinet with cell solution ...

Страница 87: ...flow rate set in the BRX DO Control menu For MCV Inoculation preferred method see Section 8 2 5 1 For FBR Inoculation see Section 8 2 5 2 4 Auto sequence FBRX DO Control Loop is in Program Manual mode white pointing finger Recirculation pump runs at Cell Attachment Flow Rate NOTE If the attachment rate needs to be changed place the recirculation pump in Operator mode and change the setpoint The At...

Страница 88: ...st hour and every 30 minutes for the subsequent 2 hours after the system is inoculated with cells Use the system volume to calculate the total number of cells attached as compared to starting cell number to determine the percentage of cells that have been depleted from the media Alternatively the concentration of cells in the system can be used to determine the attachment kinetics Inoculation is n...

Страница 89: ...tpoints Air MFC Flow Rate Stabilization Delay Press to confirm 2 Connect Inoculation bottle open and close respective clamps Connect Inoculation bottle to the FBR inoculation port Open clamp on the tubing line to the FBR Inoculation port Close the clamp on the recirculation line Return media line Return media line Recirculation line FBR Inoculation port Return media line Return media line FBR Inoc...

Страница 90: ...er all inoculum is injected close the clamp on the FBR inoculation line Open clamp on the recirculation line 6 All clamps must be manually returned to their original positions before confirming prompt to start recirculation Once the prompt is acknowledged the system begins the auto sequence steps The recirculation pump starts and the Cell Attachment Timer begins Return to Section 8 2 5 to sample t...

Страница 91: ...e pump cover Twist the gray lever to open and close the pump head TIP Make sure tubing in the pump head is centered between the V groove clamp This illustration shows the base bottle set up to automatically add base to the MCV 3 Prime Base tubing in Manual mode On the HMI select the base pump icon Press Program to run in Manual mode Press the Operator icon Stop the pump immediately before base ent...

Страница 92: ...ttle via AseptiQuik G connections on the media addition line Install the media waste bottle via AseptiQuik connections on the media waste line Press when all manual operations are complete 5 A prompt will remind the user to perform tasks to drain media from the FBR Exit the task prompt press X Open the valve on the return media line Manually jog pump in reverse to empty the bioreactor to a desired...

Страница 93: ...tion Concentration of DNA in media µg mL Depending on the transfection method the amount of transfection reagent in media may need to be adjusted Use endotoxin free DNA Adjust the volume of transfection reagent added to the MCV depending on protocol 1 Prepare transfection reagent Fill a bottle with transfection complex in the biosafety cabinet The bottle should have an AseptiQuik G connection 2 Co...

Страница 94: ...uration are completed 8 Once Stabilization Time has elapsed the system will return loops to Automatic control starting at stabilization setpoints and the user will be prompted that Transfection is complete 9 Press to acknowledge that Transfection is completed 8 2 8 Transfection Using Feed Pump This procedure is an alternative to injecting transfection reagent directly into the MCV by using the fee...

Страница 95: ...ugh they are typically run after Media Maintenance which puts all control loops in Program Auto mode To start an Auxiliary Phase during an active Sequence Phase perform the following steps 1 Select the Auxiliary Phase 2 Enter the Auxiliary Phase setpoints 3 Press to start the Auxiliary Phase The system automatically places the active Sequence Phase on Hold to allow the selected Auxiliary Phase to ...

Страница 96: ...ectors for substrate sampling 1 Select Auxiliary Phases Select Substrate Sampling 2 Enter setpoints to minimize perturbations of culture conditions Air MFC Setpoint Agitator Setpoint Stabilization Delay Duration e g 10 mins This is the time the parameters control loops are held at pre sampling setpoints before returning the loops to Automatic control Recirc Pump Ramp Duration This is the duration ...

Страница 97: ...rom the reactor Follow prompt to disconnect the bioreactor 6 Remove the FBR Disconnect the bioreactor temperature sensor Move the tray to access the Lynx connectors Disconnect the Lynx connectors and place them on the tray Transfer the bioreactor and Lynx connectors in the tray from the system to the biosafety cabinet 7 Substrate sampling in the biosafety cabinet Turn the twist clamp on the biorea...

Страница 98: ...esh segment from under the top end cap using sterile forceps or tweezers NOTE Each of the 3 sampling layers has 6 equally sized segments Place the removed segment in a sterile container Return the top end cap on the bioreactor vessel aligning the cap with the guidepost in the bioreactor Reattach the sanitary clamp to the reassembled bioreactor vessel and tighten the quick clamp to secure in place ...

Страница 99: ...onnected On the HMI press to confirm that the bioreactor is reconnected The process resumes Both timers start counting down NOTE During Stabilization Delay the system is maintained in Program Manual mode at the loop setpoint just prior to starting Substrate Sampling to allow FBR culture conditions to stabilize Meanwhile the recirculation pump ramps from the minimum flow rate to the pre sampling se...

Страница 100: ...Phase Press to acknowledge prompt that Media Removal is complete 8 3 3 Media Addition Phase This procedure shows how to add extra media to the MCV The media addition bottle can be set up in advance up to 24 hours and allowed to sit at room temperature This allows for remote addition if desired NOTE The user is responsible for visually monitoring the media level in the MCV to ensure levels are not ...

Страница 101: ...ts Media Addition Volume to prevent sending a volume to the MCV which exceeds the fill limit The window with flashing red and white fields warns the user of the error 4 Press to acknowledge prompt that Media Addition is complete 8 3 4 Bolus Addition This phase is used to add feeds bolus to the MCV at user defined instances NOTE The user is responsible for visually monitoring the media level in the...

Страница 102: ...w the expected level over time 1 Select Auxiliary Phases Select Perfusion 2 In this example for each cycle 80 mL of media will be removed from the MCV into the Media Waste bottle at 70 mL min flow rate Then 80 mL of fresh media will be added to the MCV at 70 mL min flow rate Enter setpoints Media Pump Addition SP e g 70 mL min Media Pump Removal SP e g 70 mL min Media Volume e g 80 mL It is recomm...

Страница 103: ...s responsible for visually monitoring the media level in the MCV to ensure levels are not unintentionally rising above or sinking below the expected level over time 1 Select Auxiliary Phases Select Media Dilution 2 Enter setpoints Media Pump Addition Flow Setpoint e g 50 mL min Media Pump Removal Flow Setpoint e g 50 mL min Media Volume e g 100 mL Number of Cycles e g 20 Cycle OFF Duration min Pre...

Страница 104: ...with the black clip in top valve position farthest away from the instrument Figure 8 7 Harvest Kit connection to the bioreactor 1 Scan harvest kit consumables Scan bar codes on the Harvest Kit sterile packaging and remove the consumables from the bag 2 Fill the Harvest Wash bottle with DPBS and Harvest Solution bottle with Accutase or other proteolytic reagent in aseptic conditions 3 While the har...

Страница 105: ...lue pinch valve The black tubing line connects to the Waste bottle 5 Green pinch valve tubing load Locate the 3 way tubing line with green clips At one end is the AseptiQuik connection to the bioreactor inlet At the other end are 2 tubing lines that connect to the Harvest collection and Harvest Solution Install the green white tubing line into the green pinch valve Aseptically connect the Harvest ...

Страница 106: ...install in the yellow pinch valve The yellow white line connects to the Harvest Wash Locate the yellow black tubing line and install in the yellow pinch valve The yellow black line connects to Harvest Solution 7 Flush bottle air line connection Connect the flush bottle air line to the harvest pressure port 8 AseptiQuik DC Connection The Harvest Collection bottle has the AseptiQuik DC connector mat...

Страница 107: ...hows a close up of the AseptiQuik DC connector insert which mates with the body 9 On the HMI select Harvest Phases Select Harvest Prep 10 Press to run Harvest Preparation Phase All equipment module controls are automatically stopped MCV heater FBR heater and pumps are turned off 11 Fluidically isolate the bioreactor Clamp the bioreactor inlet and return tubing lines to the MCV ...

Страница 108: ...arvest Kit to the bioreactor inlet and outlet Locate the blue tubing line Aseptically connect the tubing line to the bioreactor outlet line using the AseptiQuik G connectors Locate the yellow tubing line Aseptically connect the tubing line to the bioreactor inlet line using the AseptiQuik G connectors Make sure the AseptiQuik connectors are tightly mated to an audible click 13 Open harvest lines O...

Страница 109: ... Harvest Phase This Harvest Wash Phase is used to wash residual spent media from the FBR by pumping solution from the Harvest Wash bottle from the bottom to the top of the bioreactor to the waste bottle Figure 8 9 1 On Harvest Phases select Harvest Wash 2 Enter setpoints Recirculation Pump Flow Setpoint Wash Volume Press to run Harvest Wash Phase NOTE The phase limits Wash Volume to prevent wash b...

Страница 110: ... Harvest Wash Flow Rate until totalizer reaches Harvest Wash Volume 4 User shall acknowledge the Harvest Wash Phase completion prompt 8 4 3 Cell Release This Harvest Phase is used to release cells from the FBR by circulating solution from the Harvest Solution bottle through the bioreactor through the Harvest Flush bottle and back to the Harvest Solution bottle Figure 8 10 Figure 8 10 Cell Release ...

Страница 111: ...timer elapses recirculation pump stops 4 User shall acknowledge the Cell Release Phase completion prompt 8 4 4 Cell Removal This Harvest Phase is used to transport the released cells and solution from the Harvest Flush bottle and FBR into the Harvest Collection bottle by the release of the solution in the pressurized system via the pressure control valve Figure 8 11 1 On Harvest Phases select Cell...

Страница 112: ...timer When Cell Removal Duration timer elapses stops automated control of pressure control valve and de energizes 3 way valve for harvest pressure to vent 4 User shall acknowledge the Cell Removal Phase completion prompt 8 4 5 Final Rinse This Harvest Phase runs Harvest Wash solution through the system to collect any remaining cells in the FBR and corresponding tubing This additional cell removal ...

Страница 113: ...ms listed in Table 8 4 Table 8 4 Consumables and items required Cat No Qty Description 11440 3 1L Erlenmeyer flask with dip tube 6982 1 or 2 3L Erlenmeyer flask with AseptiQuik G connector Luer syringes for sampling Collection vials or tubes for samples Optional AseptiQuik G T adaptor if multiple collection bottles is desired and welding is not available Optional AseptiQuik S to 1 8 flexible tubin...

Страница 114: ...ion pump must be running to change the temperature otherwise the interlock prevents any changes 4 Set the recirculation pump at 500 mL min counterclockwise forward flow Set pH to 8 0 to allow for the addition of sodium bicarbonate to maintain pH Perfuse the bioreactor for 4 hours or until lysis is complete 5 Drain lysis buffer from the bioreactor to the MCV Open the air valve on media return line ...

Страница 115: ...eed to be attached earlier in the run to have appropriate connections Use an unused media addition line and connect via AseptiQuik connector 2 Perfuse salt solution through the bioreactor On the HMI go to Bolus Addition press to add High Salt Wash Or on the HMI go to Media Addition and set parameters to add High Salt Wash to the MCV In Operator mode set the recirculation pump at 500 mL min counter...

Страница 116: ...es Contact Corning for service All mass flow controllers Annual calibration See procedure in Section 8 2 3 1 All transmitters Annual calibration All pumps Annual calibration See procedure in Section 8 1 Inspect every 3 months for accumulation of tubing particulate around pump head particularly the recirculation pump Remove particulate as appropriate Table 9 2 Replaceable parts list Description Man...

Страница 117: ...s on the back of the instrument 3 Remove old gas filter Apply pressure against the metal tab below the gas connector to release the adapter and gas filter While the tab is depressed pull out the filter with the adapter 4 Separate connector from gas filter Once disconnected from the instrument remove the gas tubing connector by depressing the orange tab and pulling on the connector Remove the push ...

Страница 118: ...ufacturing Cat No PFF10000 on the Corning Ascent FBR instrument Figure 9 2 Figure 9 2 The 5 enclosure vents and their locations 1 Make sure the instrument is powered off 2 Remove the vent cover Use your fingers to pull open the top of the vent cover If the cover is hard to pull open a flathead screwdriver may be used as a wedge to pry the cover off Care should be taken to prevent damage or scratch...

Страница 119: ... vent opening Slide in the vent cover bottom 5 Close the vent cover Gently press the vent cover until an audible pop This indicates that the cover is secure in position 6 When the vent cover is secured in place there should be no gap between the vent cover and the enclosure This procedure is complete ...

Страница 120: ...vice team will help arrange local service where available coordinate a return authorization number or provide a return authorization number and shipping instructions Products received without proper authorization will be returned All items returned for service should be sent postage prepaid in the original packaging or other suitable carton padded to avoid damage Corning will not be responsible fo...

Страница 121: ...ndent Gains Form The algorithm in this form each algorithm term proportional integral and derivative has a separate gain Changing one gain affects only that term and not any of the others Where CV Control value E Error in percent of span Δt Update time in seconds used by the loop Kp Proportional gain K1 Integral gain in min 1 NOTE A larger value of K1 causes a faster integral response KD Derivativ...

Страница 122: ... modules Harvest Wash Yellow blue and green pinch valves harvest pressure valve gas shutoff valve and recirculation pump Cell Release Yellow blue and green pinch valves harvest pressure valve gas shutoff valve pressure control valve and recirculation pump Cell Removal Yellow blue and green pinch valves harvest pressure valve gas shutoff valve and pressure control valve Final Rinse Yellow blue and ...

Страница 123: ...to fully actuate Prompts user to verify that orange and red pinch valves are fully actuated If the user selects Yes proceeds to the next step If the user selects No repeats valve actuation steps once User inspects pinch valves and selects prompt response De energize orange and red pinch valves Turn exhaust heater on Initialize phase button is grayed out Prompt operator that system setup is complet...

Страница 124: ... Temperature C FBR Temperature C Air MFC setpoint SCCM O2 MFC Setpoint SCCM CO2 MFC Setpoint SCCM Stabilization Duration min User enters setpoints Agitator Setpoint 100 rpm Recirculation Pump Setpoint 120 mL min MCV Temperature 38 C FBR Temperature 37 C Air MFC setpoint 95 SCCM O2 MFC setpoint 0 SCCM CO2 MFC Flow Rate 5 SCCM N2 MFC Flow Rate 0 SCCM Stabilization Duration 120 min User selects the p...

Страница 125: ...wing setpoints Cell Attachment Flow Rate mL min Seeding volume mL Inoculation Cell Attachment Duration min Recirculation Pump Ramp Time min User enters setpoints and selects the play button Start Inoculation Timer Prompts user to select MCV Inoculation or FBR Inoculation If MCV Inoculation is selected see Option A for MCV Inoculation If FBR Inoculation is selected see Option B for FBR Inoculation ...

Страница 126: ...O Starts Stabilization Delay Duration timer and MCV DO and Gas equipment module control with output held at value recorded FBR Inoculation When timer has elapsed resumes normal automated control of equipment module setpoint FBR DO When the Cell Attachment Duration has elapsed ramps recirculation pump from the Inoculation Cell Attachment Flow Rate to the minimum FBR DO equipment module output CV 0 ...

Страница 127: ...mpts user to manually jog recirculation pump in reverse to empty the bioreactor to the desired level and to confirm when complete User manually jogs pump in reverse to empty the bioreactor to the desired level and acknowledges prompt when complete Starts media pump counterclockwise at the Removal Flow Rate setpoint until Media Pump Totalizer reaches Media Removal Volume setpoint Subtracts the Remo...

Страница 128: ...n Delay Duration timer has elapsed notifies the user that Media Exchange was completed User confirms that Media Exchange is complete N A User Action System Auto Sequence Case B MCV Volume is 2 75L and the Media Removal Volume is FBR Loop Volume Starts media pump counterclockwise at the Removal Flow Rate setpoint until the Media Pump Totalizer reaches Media Removal Volume setpoint Subtracts the Rem...

Страница 129: ...ated control of equipment module setpoint If recorded output is 50 Starts equipment module pH control with its output held at the value recorded until Stabilization timer has elapsed When timer has elapsed resumes normal automated control of equipment module setpoint When Stabilization Delay Duration timer has elapsed notifies the user that Media Exchange was completed User confirms that Media Exc...

Страница 130: ... media addition User updates MCV Volume if needed De energizes the Red pinch valve Restarts equipment module loops as follows MCV DO Starts Stabilization Delay Duration timer and MCV DO and Gas control equipment module with output at value recorded earlier When timer has elapsed resumes normal automated control of equipment module setpoints Bioreactor DO When Stabilization Delay Duration timer fro...

Страница 131: ...n Delay Duration timer and MCV DO and Gas control equipment module with output held at value recorded When stabilization delay timer has elapsed resumes normal automated control of equipment module setpoints Bioreactor DO Starts the Bioreactor DO equipment module by ramping the Recirculation Pump from the Bioreactor DO Controller minimum flow rate to the output recorded The time to ramp is set by ...

Страница 132: ...lization Delay Duration timer and MCV DO and Gas control equipment module with output held at value recorded When timer has elapsed resumes normal automated control of equipment module setpoints FBR DO Starts the FBR DO equipment module by ramping the recirculation pump from the bioreactor DO controller minimum flow rate to the recorded output The time to ramp is set by the Recirculation Pump Ramp...

Страница 133: ...ddition Volume to the Current MCV Volume and the Current Total System Volume Red pinch valve de energizes Starts Media Addition OFF Duration timer and proceeds to the next step when elapsed For setpoint defined number of cycles Repeat steps Red pinch valve is energized to open to media container through Start Media Addition OFF Duration timer Proceeds to the next step when timer has elapsed Prompt...

Страница 134: ...ion notification N A Media Removal User Action System Auto Sequence User initiates Media Removal at the HMI Phase window pops up for the user to enter the following setpoints Media Removal Flow Rate mL min Media Removal Volume mL User enters setpoints and selects the play button All current phase automated controls remain active during media removal Orange and red pinch valves are de energized Res...

Страница 135: ...Auto Sequence User initiates Media Dilution at the HMI Phase window pops up for the user to enter the following setpoints Dilution Addition Flow Rate mL min Dilution Removal Flow Rate mL min Dilution Volume mL Number of Dilution Cycles Dilution Cycle OFF Duration min User enters setpoints and selects the play button All current phase automated controls remain active during perfusion Orange pinch v...

Страница 136: ...User initiates Harvest Wash at the HMI Prompts user to enter the following setpoints Harvest Wash Flow Rate mL min Harvest Wash Volume mL User enters setpoints and selects the Play button Yellow pinch valve is energized to open to Harvest Wash Blue pinch valve is de energized to open to Harvest Waste Green pinch valve is de energized to open to Harvest Solution Harvest Pressure valve is de energiz...

Страница 137: ...he phase is complete User acknowledges Cell Removal Phase completion N A Final Rinse Phase After this cycle is complete the user will run Cell Removal Phase to collect all rinse solution in Cell Harvest bottle User Action System Auto Sequence User initiates Final Rinse at the HMI Prompts user to enter the following setpoints Final Rinse Pump Flow Rate mL min Final Rinse Volume mL User enters setpo...

Страница 138: ... red valves 3 Connect the MCV connections to the interface panel Agitator motor MCV sparge and pressure lines Exhaust filter heating blanket MCV temperature sensor Make sure media return line valve is turned off Set up the sensor tray Tilt sensors sideways prior to install MCV DO sensor Blue clip Close clamp on unused AseptiQuik G connection at the sensor tray Check that clamps on sensor tray are ...

Страница 139: ...ation DO sensor calibration Enter a new reference value typically 100 of air saturation pH sensor calibration Take a media sample and measure pH level offline Enter the new pH Reference Value Temperature sensor calibration MCV temperature offset is typically 3 C FBR temperature offset is typically 0 5 C 3 Select Media Maintenance phase Enter parameters Section 8 2 4 MCV TIC Temp setpoint 0 to 70 C...

Страница 140: ...afety cabinet Perform substrate sampling Reconnect the Lynx connectors after sampling if using Reconnect the FBR temperature sensor 7 Select Media Exchange phase Media shall be at the correct temperature before addition otherwise add cold media slowly Section 8 2 6 Remove Media Reservoir from connector Connect via AseptiQuik connections to the new Media Addition bottle Connect via AseptiQuik conne...

Страница 141: ...s and pumps are turned off 6 Clamp the bioreactor inlet and return tubing lines to the MCV 7 Load yellow tubing line in the recirculation pump 8 Connect the Harvest kit to the bioreactor inlet and outlet Mate AseptiQuik connectors to an audible click 9 Remove both clamps on the Harvest lines Harvest Collection Table A4 4 Harvest collection check list Label Harvest collection check list Check Refer...

Страница 142: ...cified all products are for research use or general laboratory use only Not intended for use in diagnostic or therapeutic procedures Not for use in humans These products are not intended to mitigate the presence of microorganisms on surfaces or in the environment where such organisms can be deleterious to humans or the environment Corning Life Sciences makes no claims regarding the performance of ...

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