CORNING Ascent 6970 User manual

Corning®Ascent™Fixed Bed Reactor
Process Development System
Instruction Manual
Catalog Number: 6970

CORNING ASCENT FBR PD SYSTEM
Table of Contents
1.0 Safety........................................................................ 1
1.1 Personal Protective Equipment (PPE)................................. 1
1.2 Equipment Safety ................................................................... 1
2.0 Introduction ............................................................. 2
3.0 Site Requirements ................................................... 3
4.0 Set Up and Installation............................................ 4
4.1 Power, Air, and Gas Connections......................................... 4
5.0 Instrument Overview .............................................. 5
5.1 Pre-sterilized Consumables.................................................. 5
5.1.1 Media Addition and Media Waste Bottles ............ 5
5.1.2 Media Conditioning Vessel (MCV) ........................... 5
5.1.3 Sensor Tray ..................................................................... 7
5.1.4 Inoculum, Base, and Feed Bottles ........................... 8
5.1.5 FBR with Sterile Connectors...................................... 8
5.1.6 Harvest Kit ..................................................................... 9
5.1.7 Sterile Connectors......................................................11
5.2 Interface Panel.......................................................................11
5.2.1 Agitator Motor............................................................12
5.2.2 Exhaust Filter Heater.................................................12
5.2.3 Pressure Sensor...........................................................13
5.2.4 MCV Sparge .................................................................13
5.2.5 MCV Temperature Sensor.........................................13
5.2.6 MCV and FBR Dissolved Oxygen (DO) Sensors...14
5.2.7 MCV pH Sensor...........................................................14
5.3 Pumps ......................................................................................14
5.4 Pinch Valves............................................................................15
5.4.1 Pinch Valve Positions.................................................16
5.5 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 .........................................................22
6.2.4 Transfection .................................................................23
6.3 Auxiliary Phases .....................................................................23
6.3.1 Substrate Sampling ...................................................23
6.3.2 Media Addition ...........................................................23
6.3.3 Bolus (or Feed) Addition ...........................................23
6.3.4 Media Removal ...........................................................24
6.3.5 Perfusion or Media Replacement...........................24
6.3.6 Media Dilution............................................................24
6.4 Harvest .....................................................................................24
6.4.1 Harvest Preparation...................................................25
6.4.2 Harvest Wash ..............................................................25
6.4.3 Cell Release ..................................................................25
6.4.4 Cell Removal ................................................................26
6.4.5 Final Rinse ....................................................................27
7.0 Human-Machine Interface (HMI)..........................27
7.1 Main Screen Overview.........................................................27
7.3 Access Levels and User Modes...........................................31
7.4 System Screen........................................................................32
7.5 Phase Status, Abort, and Global Reset ............................33
7.6 Media, Harvest, and Auxiliary Phases .............................34
7.6.1 Navigating Sequence and Auxiliary Phase
Faceplates.....................................................................37

Instruction Manual
7.7 Calibration ..............................................................................37
7.8 Equipment Control Faceplates ..........................................38
7.8.1 Equipment Parameter Control Faceplates...........39
7.8.2 Equipment Totalizer Faceplate ...............................40
7.8.3 Proportional, Integral, Derivative (PID) Loop
Faceplate ......................................................................40
7.9 Control Loop Setup ...............................................................42
7.9.1 MCV DO Control .........................................................42
7.9.1.1 MCV DO Control Loop..................................43
Configuration Examples
7.9.2 MCV pH Control..........................................................44
7.9.3 Bioreactor DO Control...............................................46
7.10 Accessing Trend Properties...............................................46
7.10.1 Create a New Time Period .....................................47
7.10.2 Tag List Features.......................................................48
7.10.3 Display Trace..............................................................49
7.10.4 Templates...................................................................51
7.10.4.1 How to Create a Template....................51
7.10.4.2 Apply an Existing Template..................51
7.10.4.3 Save Changes to Existing Template...52
7.10.4.4 Add Tags to Template ............................52
7.10.4.5 Remove Template Tags..........................54
7.10.4.6 Zoom and Navigate Data .....................54
7.10.4.7 Trend Properties......................................55
7.11 Process Data.........................................................................55
7.11.2 Data File Entry........................................................56
7.11.4 USB Save..................................................................57
7.12 Diagnostics...........................................................................58
7.13 VPN Enabled.........................................................................58
7.13.1 Remote Access........................................................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 Inoculation (Preferred Option).......84
8.2.5.2 FBR Inoculation ............................................85
8.2.5.3 Adding Base...................................................87
8.2.6 Media Exchange Phase .............................................87
8.2.7 Transfection Phase (Process-dependent) .............89
8.2.8 Transfection (Using Feed Pump).............................90
8.3 Auxiliary Phases ....................................................................91
8.3.1 Substrate Sampling ...................................................92
8.3.2 Media Removal Phase...............................................96
8.3.3 Media Addition Phase...............................................96
8.3.4 Bolus Addition.............................................................97
8.3.5 Perfusion.......................................................................98
8.3.6 Media Dilution Phase (Process-dependent)........99
8.4 Harvest Phases ......................................................................99
8.4.1 Harvest Preparation................................................ 100
8.4.2 Harvest Wash ........................................................... 105
8.4.3 Cell Release ............................................................... 106
8.4.4 Cell Removal ............................................................. 107
8.4.5 Final Rinse ................................................................. 108
8.5 In Situ Lysis Manual Protocol........................................... 109
8.5.1 Remove Cell Culture Media .................................. 109
8.5.2 Lysis............................................................................. 110
8.5.3 PBS Wash................................................................... 110
8.5.4 High Salt Wash (Optional) .................................... 111
8.6 Shutdown Procedure......................................................... 111

CORNING ASCENT FBR PD SYSTEM
9.0 Preventative Maintenance..................................112
9.1 Recommended List ............................................................ 112
9.2 Routine Screen Cleaning .................................................. 112
9.3 Gas Filter Replacement Procedure................................. 113
9.4 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

1
Instruction Manual
1.0 Safety
This section describes how to safely set up and use the Corning® Ascent™ Fixed Bed Reactor Process Development (FBR PD) System.
1.1 Personal Protective Equipment (PPE)
Non-toxic media is handled during seeding and harvest. Basic PPE shall include:
• Safety eye wear
• Laboratory coat
• Examination gloves when handling media
• Face shield particularly during Harvest protocols
1.2 Equipment Safety
Table 1-1. Warning 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.
Heavy object symbol. To avoid injury, lifting the Ascent instrument requires two
or more people as per OSHA standard.
Read and fully understand the Instruction Manual before using the instrument.
Failure to do so could result in injury.

2CORNING ASCENT FBR PD SYSTEM
Figure 2-2. Bottle compartment with bar code scanner (see Table 2-1).
Figure 2-1. Corning Ascent FBR PD system
(see Table 2-1).
2.0 Introduction
The Corning® Ascent™ FBR PD System consists of a system controller and single-use, pre-sterilized components including a bioreactor, a
Media Conditioning Vessel (MCV), and other consumables, such as tubing, connectors, probes, in-line sensors, and bottles (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 temperature. At harvest,
cells are released or lysed in situ from the bioreactor at the user’s discretion. A Human-Machine Interface (HMI) allows the user flexibility to
operate the system in manual or automated modes.

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Instruction Manual
Table 2-1. Summary table of the main components.
Label Component Reference
1Human-Machine Interface (HMI) Section 7.0
2Media Conditioning Vessel (MCV) Section 5.1.2
3Fixed Bed Reactor (FBR) with sterile connectors tray Section 5.1.5
4Bioreactor heated nest Section 5.5
5Recirculation pump Section 5.3
6Media pump Section 5.3
7Pinch valve(s) Section 5.4
8Interface panel Section 5.2
9Inoculum, base, and feed bottles rack Section 5.1.4
10 Media Addition and Media Waste bottles Section 5.1.1
11 Bar code scanner Section 5.6
3.0 Site Requirements
Table 3-1 lists the site requirements to set up the Corning® Ascent™ FBR PD instrument.
Table 3-1. Site requirements.
Type Description
Operating Conditions The instrument is designed for use under standard operating conditions per UL 61010-1 where:
• 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 76.2 x 68.58 cm).
The workbench shall be sized to allow additional consumables, if required.
System Weight Base weight (without consumables) is approximately 186 lbs.
Power Requirements Cat. No. 6970, 120 VAC/60 Hz/1-Phase 527 W
Cat. No. 6991, 230 VAC/50 Hz/1-Phase 595 W
One 20 amp. dedicated outlet is required
An uninterruptible power supply (UPS) and a surge protector are recommended.
Gas Connection
and Type
All calibrated mass flow controllers (MFCs) are rated for 30 to 35 psig of their respective gases.
Required connection type is 1/4" (6 mm) OD semi-rigid flexible tubing for inlet push connectors.
Communication EtherNet/IP

4CORNING ASCENT FBR PD SYSTEM
Figure 4-2. Control system power On/Off button,
USB port, and EtherNet/IP port.
Figure 4-3. Pressure port connection at harvest.
Figure 4-1. Power, air, and gas connections at the
back of the instrument.
4.0 Set Up and Installation
This section describes how to connect and power up 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 starting a run. A pressure port (Figure 4-3) along the instrument
base left corner is only connected at harvest.

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Instruction Manual
5.0 Instrument Overview
This section describes the Corning® Ascent™ FBR instrument, which includes the pre-sterilized consumables and the system controller.
5.1 Pre-sterilized Consumables
Table 5-1 is a list of the pre-sterilized, single-use consumables that shall be properly discarded as biohazardous waste, as mandated by
local and state laws. Consumables shall not be reused.
Table 5-1. Consumables overview.
Consumable Illustration Reference
Media Conditioning Vessel (MCV) with sensor tray Figure 5-1 Sections 5.1.2 and 5.1.3
Media Addition bottle Figure 2-2 Section 5.1.1
Media Waste bottle Figure 2-2 Section 5.1.1
Inoculum bottle Figure 5-5 Section 5.1.4
Base bottle Figure 5-5 Section 5.1.4
Feed bottle Figure 5-5 Section 5.1.4
Fixed Bed Reactor (FBR) and sterile connectors Figures 5-6 and 5-7 Section 5.1.5
Harvest Wash bottle Figure 5-10 Section 5.1.6
Harvest Solution bottle Figure 5-10 Section 5.1.6
Harvest Collection Bottle Figure 5-10 Section 5.1.6
Flush bottle Figure 5-10 Section 5.1.6
Waste bottle Figure 5-10 Section 5.1.6
Figure 5-1. Media recirculation and return
lines via the Lynx® connector, and sterile
connectors to 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 media exchange, spent media is discarded into the media waste bottle which also resides in the same housing on the side of the
instrument. It is connected via parallel tubing that runs from the MCV. Media waste is removed through the tubing by the media pump.
More than one media waste bottle can be connected as needed.
There are two pinch valves, the red and orange pinch valves, that work in conjunction to direct either fresh media to the MCV or remove
spent media from the MCV (see Section 5.4 for details).
5.1.2 Media Conditioning Vessel (MCV)
The media pump flows fresh media to the MCV, and the recirculation pump perfuses media through the bioreactor. The MCV sits in a
heated nest while its contents are stirred at a programmed speed to maintain homogeneity, and to maintain desired dissolved oxygen
(DO) levels in the media during cell culture. Figure 5-1 illustrates an overview of the tubing connections between the MCV and bioreactor
via the Lynx® connectors.

6CORNING ASCENT FBR PD SYSTEM
Table 5-2. Tubing and ports on the MCV headplate.
Label Description IN OUT
T1 Not used.
T2 Adds and removes media from the MCV.  
T3 Channels MCV media through the sensor tray and sterile connections to the
bioreactor. The tubing has a sensor that monitors the MCV temperature. 
T4
Returns media from the bioreactor. There are two features on the line:
• Media sampling port.
• Valve is manually opened when 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 port. 
p2 Media sampling Luer-Lock port. 
Figure 5-2. MCV headplate connections
for media maintenance, transfection,
inoculation, and media sampling
(see Table 5-2).
Figure 5-2 shows the MCV headplate connections, tubing lines, and ports.

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Instruction Manual
Inside the MCV are the following (Figure 5-3):
• An agitator shaft with a rotating 6-blade Rushton impeller that facilitates homogenous media mixing
• A dip tube connected to the gas/sparging line
• A dip tube for media addition, media removal, and media return
• A dip tube for media recirculation from MCV to the bioreactor
• A port segment for return media from the bioreactor (not visible in Figure 5-3)
5.1.3 Sensor Tray
The sensor tray fits between the MCV and the recirculation pump (Figure 5-4) to properly position the sensors and recirculation 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 MCV and FBR dissolved oxygen levels, and MCV pH
Figure 5-3. Inside the MCV is a
powered agitator; a gas/sparge line
dip tube; media addition, removal
and return dip tube; and the MCV
media recirculation dip tube.
Figure 5-4. The sensor tray contains
media tubing with in-line pH and
DO sensors that are clamped to
its surface for monitoring media
conditions.

8CORNING ASCENT FBR PD SYSTEM
Table 5-3. Base, feed, and inoculum bottles descriptions.
Bottle Description
Base (or alkali) This bottle contains a basic solution to adjust cell culture media pH inside the MCV. The
Ascent system is designed to automatically maintain the MCV cell culture media pH by
adding base solution via the base pump and controlling pump flow with user-defined
setpoints. Adding a base solution during cell growth neutralizes excessive concentration of
H+ ions and hence prevents the media from being too acidic. An example of a base solution is
7.5% sodium bicarbonate solution.
Feed (or bolus) This bottle contains concentrated supplement(s) that, when added to the media, can increase
nutrient concentrations during cell culture. Feed is pumped into the MCV at a programmed
rate via the feed pump.
As an option, nutrients such as glucose (45%) and L-glutamine (200 mm) can be added in
bulk via the feed line to increase concentrations.
Inoculum (or cell
suspension)
This bottle contains a concentrated cell suspension in media that is used to seed the
bioreactor. Seeding can be done through the 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® connectors provide users the added benefit of aseptically connecting and disconnecting the FBR up to six times for manual
manipulations enabling substrate sampling. The portable tray allows users the convenience of moving the disconnected FBR and Lynx®
connectors to a biosafety cabinet. The AseptiQuik® G is a single-use connector and cannot be disconnected or reconnected.
5.1.4 Inoculum, Base, and Feed Bottles
Inoculum is manually added during an experiment. The user selects one of two AseptiQuik® G connectors on the MCV headplate (see
Figure 5-2). Base is automatically added as part of the pH control loop. Base and feed lines have respective pumps that require set up prior
to use. Figure 5-5 shows a rack for base, feed, and inoculum bottles. Refer to Table 5-3 for descriptions.
Figure 5-5. Base and feed bottles are connected to their respective
pumps during an auto-cycle. Inoculum is manually introduced.

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Instruction Manual
The bioreactor has a sanitary twist clamp that allows easy access to the segmented sampling mesh located below (Figure 5-8). The 3
sampling mesh disks (Figure 5-9) are divided into six segments, which are designed for easy removal. Segments may be removed to
monitor cell growth in the FBR.
Figure 5-6. FBR and Lynx® connectors. Figure 5-7. FBR and AseptiQuik® G connectors.
Figure 5-9. A segmented sampling mesh. There
are 3 segmented sampling meshes, each with 6
segments, totaling 18 sampling segments. One
or more segments may 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 conjunction with the automated pinch valves, perfuse harvesting solutions from the harvest kit
through the bioreactor. See Table 5-4 for harvest kit descriptions.
Figure 5-10. Illustration showing the harvest kit.
The kit is only set up at harvest.
Figure 5-8. The growth mesh
stack and sampling mesh
layers are removable from
the bioreactor vessel.

10 CORNING ASCENT FBR PD SYSTEM
Figure 5-11. Harvest Solution flow
layout with Lynx® connectors.
Harvest tubing lines are connected
via AseptiQuik® G connectors. Tubing
clamps close Lynx® connector lines to
allow wash and harvest solutions to
enter the bioreactor.
Table 5-4. Harvest kit.
Label Bottle/Vessel Description
HC Harvest Collection This bottle is used to collect cells harvested from the bioreactor. During the harvest step, harvest solution
flushes out the cells from the bioreactor, and the cells suspension is collected in the collection bottle.
HS Harvest Solution This bottle contains harvest solution such as Accutase® or a similar proteolytic cell dissociating enzyme.
For complete recovery of the harvested cells from the bioreactor, it is recommended to perform a post-harvest
bioreactor wash with DPBS solution.
HW Harvest Wash This bottle contains DPBS to remove cell culture media from the bioreactor 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 ‘flush’ cells from the
bioreactor.
There is always approximately 80-100 mL of harvest solution in the harvest flush bottle.
W Waste This bottle collects the wash solution that has passed through the bioreactor – before the proteolytic agent is
added – for disposal.
Figure 5-11 shows the harvest solution flow layout with Lynx® connectors. During harvest, the Lynx® connectors remain connected to
the MCV, but the tubing clamps are manually closed at harvest consumables setup to re-direct flow through the AseptiQuik® connectors.
Figure 5-12 shows the harvest layout on instruments using AseptiQuik® G connections on the MCV recirculation loop.
Figure 5-12. Harvest Solution flow
layout with single-use AseptiQuik® G
connectors.

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Instruction Manual
5.1.7 Sterile Connectors
Table 5-5 and Table 5-6 list the sterile connectors used at cell culture and harvest.
Table 5-5. Sterile connections at Media fill, Prime, and Conditioning set up.
Lynx® CDR
Connector
AseptiQuik® G
AQG17004
AseptiQuik® S
AQS17002 Connection Purpose

(Option A)

(Option B)
MCV to bioreactor recirculation line Media perfusion

(Option A)

(Option B)
Bioreactor return media line Media perfusion
Media addition bottle 1 to MCV Media fill/addition
Media addition bottle 2 to MCV Media fill/addition
MCV to media waste bottle 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
AQCDC22004 Connection Purpose
Bioreactor to harvest collection bottle Harvest collection
Harvest line to bioreactor inlet Harvest process
Harvest line to bioreactor outlet Harvest process
5.2 Interface Panel
Figure 5-13 shows the interface panel for various sensors and components connections (see Table 5-7 for details).
Figure 5-13. Interface panel ports.

12 CORNING ASCENT FBR PD SYSTEM
5.2.1 Agitator Motor
The motor driven Rushton impeller uniformly mixes the media in the MCV. The magnetically coupled agitator motor is manually mounted
on the MCV headplate (Figure 5-14) to drive the agitator shaft (Figure 5-3). Agitation speed can be controlled manually or automatically to
maintain proper level of media oxygenation in the MCV.
The recommended agitation operational range is 0 to 400 rpm, and the speed can be set up to 500 rpm via the HMI.
NOTE: It is not recommended that the agitator motor cable be disconnected from either end as motor malfunction may occur. Motor
should be kept in the motor holder when not being used on an MCV (see Figure 5-14).
Figure 5-14. Agitator motor
connection is plugged into
the interface panel. To start or
stop the agitator, the motor is
manually mounted or removed
from the MCV headplate.
Label Interface Panel Description Reference
1Agitator motor Agitator motor cable Section 5.2.1
2Filter heater Exhaust filter heater connection Section 5.2.2
3MCV pressure In-line sensor that monitors pressure on the MCV gas vent Section 5.2.3
4MCV sparge Sparging gas line in the MCV Section 5.2.4
5MCV temperature In-line sensor monitors the media temperature exiting the
MCV via the recirculation loop
Section 5.2.5
6FBR DO (DO2) Monitors dissolved oxygen levels exiting the bioreactor Section 5.2.6
7MCV pH Monitors pH levels exiting the MCV Section 5.2.7
8MCV DO (DO1) Monitors dissolved oxygen levels exiting the MCV Section 5.2.6
Table 5-7. Interface panel ports.
5.2.2 Exhaust Filter Heater
An exhaust line (Figure 5-15) prevents pressure build-up in the MCV. The exhaust line has a filter that requires heating to prevent
condensate build-up. Heating is performed via a reusable silicone heater blanket that keeps the filter dry. The heater blanket is manually
secured via buttons around the filter element before a process run so that the filter is immediately heated when gas flow begins. The
heater temperature is controlled to a fixed power output. A built-in thermostat prevents the temperature from rising above 65°C. A wire
rack keeps the exhaust line with the secured heater blanket supported.
NOTE: The vent filter heater is only turned off at system shutdown. It remains on during harvest and in situ lysis.
Figure 5-15. Heater blanket
wrapped around the exhaust
filter on the gas line.

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Instruction Manual
5.2.3 Pressure Sensor
Located below the exhaust filter is the MCV pressure sensor (Figure 5-16). It is designed to detect a low or high-pressure event within the
MCV that can occur (e.g., if the vent filter clogs because of condensate build-up or foam entry into the exhaust filter). The sensor relays
the high-pressure event to the controller. The controller responds by shutting off the gas valves and putting all loops in a safe state until
the pressure condition is cleared, or the experiment is stopped via a shutdown.
The normal operating pressure range is between -0.5 to 2.0 psi. When the pressure hits -0.5 psi, the system alarms. When the pressure
reaches +2 psi, all mass flow controllers are set to 0 sccm, and the active current phase is put on HOLD.
5.2.4 MCV Sparge
When the gas/sparge line (Figure 5-17) is plugged into the interface panel via an MPC connection, O2, air, CO2, and N2flow is regulated
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 (MFC) setpoints are zero
and opens when MFC setpoints are non-zero. The valve is normally closed when not energized (e.g., if the system is powered off). The
valve also closes when the pressure sensor (see Section 5.2.3) detects a high-pressure event.
Figure 5-16. MCV pressure sensor
monitors the exhaust line.
Figure 5-17. Gas line on the
MCV is monitored.
5.2.5 MCV Temperature Sensor
An in-line temperature sensor in the MCV media outlet monitors the temperature of the cell culture media exiting the MCV (Figure 5-18).
The in-line sensor cannot measure MCV media temperature accurately if the recirculation pump is not running.
Figure 5-18. MCV temperature
sensor monitors warmed
media flowing from the MCV
to the bioreactor.

14 CORNING ASCENT FBR PD SYSTEM
5.2.6 MCV and FBR Dissolved Oxygen (DO) Sensors
Two dissolved oxygen (DO) sensors are located on the sensor tray. The MCV DO is an in-line sensor located in the tubing section
between the MCV and bioreactor. The FBR DO sensor is also an in-line sensor located on the other side of the tubing loop between the
bioreactor and MCV (see Figure 5-19 and Figure 5-20). Both sensors depend on the recirculation pump to run seamlessly for accurate DO
measurements at the respective vessels.
Dissolved oxygen is measured in terms of % air saturation level (i.e., 100% air saturation indicates that the gases dissolved in the media
are in equilibrium with air at normal atmospheric conditions.). The user defines the MCV DO level, which is automatically maintained by
the DO cascade control loop via three different output variables: O2% in the sparging gas mixture, sparging gas flow rate, and agitator
speed. Similarly, the user defines the FBR DO outlet level, causing the recirculation pump flow rate to increase when the DO at the
bioreactor outlet drops below the setpoint.
DO sensor calibration values are provided by the manufacturer and can be read from the bar code on the MCV Consumable set or
manually entered by the user.
Figure 5-19. MCV DO sensor. Figure 5-20. FBR DO sensor.
5.2.7 MCV pH Sensor
The pH of the cell culture media in the MCV is monitored by an in-line sensor in the tubing on the sensor tray (Figure 5-21). The pH
registered range is 5.5 to 8.5 and is user-defined. It is automatically maintained via the pH controller which controls two different outputs:
the base pump flow rate and the CO2flow 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 Pumps
The Corning® Ascent™ FBR system utilizes four peristaltic pumps programmed to perform a specific range of operations (Figure 5-22). The
two large pump heads (Figure 5-23) are used for media fill and recirculation, while the two smaller pump heads (Figure 5-24) are used for
automating feed and base additions.
The large pumps work in conjunction with the pinch valves (Section 5.4: Pinch Valves) to direct media flow. Only the tubing section in
the recirculation pump is robust enough to withstand the stress from continuous pumping when perfusing the bioreactor throughout
the process run. Both the recirculation and media pumps are programmed to rotate clockwise or counterclockwise, depending on the
process phase.
The smaller pumps, feed and base, have color-coded covers to help the user load and align tubing lines with matching color clips. The feed
and base pumps are programmed in one direction, clockwise, to add liquid into the MCV.
All four pumps are calibrated prior to a run to determine the maximum flow rate at maximum output (Section 8.1: Pump Calibration). The
user sets and controls pump speed via the HMI. The tubing diameter is larger for the recirculation and media pumps than for the base and
feed pumps. For all four pumps, the tubing is locked into position via a sliding or twist lock.

15
Instruction Manual
Figure 5-22. Peristaltic pump locations. Refer to Table 5-8 for descriptions.
Label Pump Pump Rollers Direction Flow Description
P1 Media pump Clockwise (CW) Advanced (from media bottle
to MCV)
Pump speed: approximately 0.5 to 700 mL/min. (calibration-
dependent).
Adds media from the MCV (e.g., during media exchange).
Counterclockwise (CCW) Reversed (from MCV to media
bottle)
Removes media from the MCV (e.g., during media exchange)
P2 Recirculation
pump
Clockwise (CW) Advanced (from MCV to
bioreactor inlet)
Pump speed: approximately 0.5 to 700 mL/min. (calibration-
dependent).
Circulates media between the MCV and the bioreactor.
Perfuses harvest solutions through the bioreactor.
Counterclockwise (CCW) Reversed (from bioreactor inlet
to MCV)
Drains media from the bioreactor to the MCV during substrate
sampling.
P3 Feed pump Clockwise (CW) Advanced (from feed bottle to
MCV)
Pump speed: approximately 0.1 to 80 SCCM (calibration-
dependent).
Adds nutrients to the MCV.
P4 Base pump Clockwise (CW) Advanced (from base 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 5-25) that work in conjunction with the pumps to channel and direct media flow. Each pinch valve cover
color matches the colored ring around its base. The orange and red pinch valves are used for media fill, and media removal. The yellow,
blue, and green pinch valves are used at harvest.
The pinch valve color assists the user in proper loading and positioning of tubing lines. Each tubing line is differentiated by colored clips
that match the pinch valve color (Figure 5-26). All tubing lines have black and white colored clips that are inserted in the top and bottom
valve position, respectively. Pinch valves are sized to be compatible with tubing inner diameter (ID) measuring 0.25 inches, and outer
diameter (OD) measuring 0.375 inches. Refer to Table 5-9 for pinch valves descriptions and assigned colors.

16 CORNING ASCENT FBR PD SYSTEM
NOTE: The white tubing clip is always positioned next to the instrument, while the black tubing clip is on the outside.
Figure 5-25. Pinch valve locations. Refer to Table 5-9 for descriptions.
Figure 5-26. A pinch valve close-up. The cover label
shows where to align and load the tube lines based
on the tubing clip color.
5.4.1 Pinch Valve Positions
Valve positions are important for directing tubing flow when the system calls for it. Each pinch valve secures tubing in two valve positions:
top or bottom. Both top and bottom valve positions have plungers programmed to actuate – raised or retracted, depending on where
media is directed. The top valve position, 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 place when the pinch valves are de-energized. The user shall load
tubing in the bottom valve position first followed by the top valve position.
Once a phase is started on the HMI, the pinch valves are energized. The top and bottom valve plungers actuate accordingly depending on
media direction.
Figure 5-27. A de-energized pinch valve at power-up. The top valve plunger is retracted to allow uninterrupted tubing flow.
The bottom valve plunger is raised to constrict the bottom tubing.