NEW BRUNSWICK SCIENTIFIC BioFlo CelliGen 115 User manual

New Brunswick Scientific PO Box 4005 44 Talmadge Rd. Edison, 08818-4005 USA
1.800.631.5417 1.732.287.1200 bi[email protected] www.nbsc.com
Guide to Operations
BioFlo®/CelliGen®115
Benchtop Fermentor & Bioreactor
MANUAL No: M1369-0050
Revision B
June 2, 2009

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BioFlo/CelliGen 115 •M1369-0050 User’s Guide

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New Brunswick Scientific User’s Guide
INTERNATIONAL OFFICES:
BELGIUM
New Brunswick Scientific NV-SA
Stationsstraat 180/4
3110 Rotselaar
België/Belgique
Tel: [phone]
Fax: [phone]
E-mail: [email protected]
GERMANY
New Brunswick Scientific GmbH
In Der Au 14
D-72622 Nürtingen
Deutschland
Tel: [phone]
Fax: [phone]
E-mail: [email protected]
CHINA
New Brunswick Scientific Co., Inc.
A903, 904 Yin Hai Building
No. 250, Cao Xi Road
Shanghai 200235, P.R. China
Tel: [phone] or 5966
Fax: [phone]
E-mail: [email protected]
THE NETHERLANDS
New Brunswick Scientific BV
Kerkenbos 1101, 6546 BC Nijmegen
P.O Box 6826, 6503 GH Nijmegen
Nederland
Tel: [phone]
Fax: [phone]
E-mail: [email protected]
FRANCE
New Brunswick Scientific SARL
12-14, Rond Point des Champs Elysées
75008 Paris
France
Tel: [phone]
Fax: [phone]
E-mail: [email protected]
UNITED KINGDOM
New Brunswick Scientific (UK) Ltd.
17 Alban Park
St. Albans, Herts. AL4 0JJ
United Kingdom
Tel: [phone] or 0800 581331
Fax: [phone]
E-mail: [email protected]
Web: www.nbsuk.co.uk

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BioFlo/CelliGen 115 •M1369-0050 User’s Guide
WARNING!
This product is not designed to contain gases within the range of their
lower explosion limit (LEL) and their upper explosion limit (UEL).
If your process requires or produces gases, be sure to verify their LEL
and UEL concentration range (available online).
CAUTION!
This equipment must be operated as described in this manual.
If operational guidelines are not followed, equipment damage and
personal injury can occur. Please read the entire User’s Guide before
attempting to use this unit.
Do not use this equipment in a hazardous atmosphere or with hazardous
materials for which the equipment was not designed.
New Brunswick Scientific (NBS) is not responsible for any
damage to this equipment that may result from the use of an accessory
not manufactured by NBS.
WARNING!
High voltage.
Always make sure this equipment is properly grounded.

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New Brunswick Scientific User’s Guide
Copyright Notice
New Brunswick Scientific
Box 4005, 44 Talmadge Road
Edison, New Jersey 08818-4005
©Copyright 2009 New Brunswick Scientific
All Rights Reserved.
Reproduction, adaptation, or translation without prior written permission from
New Brunswick Scientific is prohibited.
Disclaimer Notice
New Brunswick Scientific reserves the right to change information in this
document without notice. Updates to information in this document reflect our
commitment to continuing product development and improvement.
Manual Conventions
#NOTE:
Notes contain essential information that deserves
special attention.
CAUTION!
Caution messages appear before procedures which, if
caution is not observed, could result in damage to the
equipment.
WARNING!
Warning messages alert you to specific procedures or
practices which, if not followed correctly, could result in
serious personal injury.
Bold Text in boldface type emphasizes key words or
phrases.
This particular Warning message, whether found in the
manual or on the unit, means HOT SURFACE–and
therefore represents a potential danger to touch.
CRUSH WARNING!
Crush Warning messages alert you to specific
procedures or practices regarding heavy objects which,
if not followed correctly, could result in serious personal
injury .

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BioFlo/CelliGen 115 •M1369-0050 User’s Guide
Every instrument manufactured by New Brunswick Scientific
is warranted to be free from defects in material and workmanship.
This apparatus with the exception of glassware, lamps and
electrodes (where supplied), is warranted for 1 year against faulty
components and assembly and our obligation under this warranty
is limited to repairing or replacing the instrument or part thereof,
which shall, within 1 year after date of shipment, prove to be
defective after our examination. This warranty does not extend
to any NBS products which have been subjected to misuse, neglect,
accident or improper installation or application; nor shall it extend
to products which have been repaired or altered outside the
NBS factory without prior authorization from
New Brunswick Scientific.
W A R R A N T Y

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New Brunswick Scientific User’s Guide
FERMENTOR/BIOREACTOR
INFORMATION SHEET
On this page, record the information for your fermentor/bioreactor and retain this for
future reference.
MODEL NUMBER: ________________________________
VOLTAGE: ________________________________
SERIAL NUMBER: ________________________________
The above information can be found on the electrical specification plate.
Purchased with the following installed options:
__________________________________________________
__________________________________________________
__________________________________________________
__________________________________________________
__________________________________________________

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BioFlo/CelliGen 115 •M1369-0050 User’s Guide
TABLE OF CONTENTS
1WARNINGS & CAUTIONS...........................................................................................1
1.1 WARNINGS .............................................................................................................. 1
1.2 CAUTIONS................................................................................................................ 2
2INSPECTION & UNPACKING OF EQUIPMENT.....................................................4
2.1 INSPECTION OF BOX(ES).............................................................................................. 4
2.2 PACKING LIST VERIFICATION...................................................................................... 4
2.3 BASIC COMPONENTS ................................................................................................... 4
3INTRODUCTION & OVERVIEW................................................................................5
3.1 SYSTEM ....................................................................................................................... 5
3.2 VESSELS ...................................................................................................................... 5
3.3 AGITATION SYSTEM .................................................................................................... 5
3.4 TEMPERATURE CONTROL ............................................................................................ 6
3.5 AERATION ................................................................................................................... 6
3.6 PHCONTROL ............................................................................................................... 6
3.7 DO CONTROL.............................................................................................................. 7
3.8 FOAM/LEVEL CONTROL .............................................................................................. 7
3.9 EXHAUST SYSTEM ....................................................................................................... 7
3.10 RECOMMENDED ACCESSORIES &SUPPLIES................................................................. 7
3.11 SUPERVISORY SOFTWARE ........................................................................................... 8
4INSTALLATION.............................................................................................................9
4.1 PHYSICAL LOCATION .................................................................................................. 9
4.2 ENVIRONMENT ............................................................................................................ 9
4.3 INSTALLING THE CONTROL CABINET......................................................................... 10
4.4 CONNECTING UTILITY CABINETS .............................................................................. 12
4.5 UTILITIES .................................................................................................................. 14
4.5.1 Electrical Requirements...................................................................................14
4.5.2 Water and Drain Connections..........................................................................15
4.5.3 Gas Connections ..............................................................................................16
4.6 **IMPORTANT WARNINGS**..................................................................................... 17
4.7 VESSEL ASSEMBLY:NON-JACKETED........................................................................ 19
4.7.1 Headplate.........................................................................................................22
4.7.2 Install Heat Blanket..........................................................................................24
4.7.3 Install Vessel in Vessel Stand...........................................................................25
4.7.4 Install Baffle (14.0L Fermentation Vessels ONLY)..........................................26
4.8 VESSEL ASSEMBLY:WATER-JACKETED ................................................................... 26
4.8.1 Install Headplate Clamping Ring.....................................................................28
4.8.2 Install Vessel on Base Plate .............................................................................28
4.8.3 Filling the Water Jacket...................................................................................29
4.8.4 Install Baffle (14.0L Fermentation Vessels ONLY)..........................................29
4.8.5 Install Impeller(s).............................................................................................30

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4.8.6 Install Cooling Coil..........................................................................................31
4.8.7 Install Sparger (3.0L, 7.5L & 14.0L Vessels)...................................................31
4.8.8 Install Harvest Tube.........................................................................................32
4.8.9 Install Sampler Tube ........................................................................................32
4.8.10 Install Thermowell............................................................................................32
4.8.11 Install Foam Probe...........................................................................................32
4.8.12 Install Foam Exhaust Tube ..............................................................................33
4.8.13 Install Level Probe(s).......................................................................................33
4.8.14 Install Addition Tube(s)....................................................................................33
4.8.15 Install pH Probe...............................................................................................33
4.8.16 Install dO2 Probe.............................................................................................35
4.8.17 Install Exhaust Condenser ...............................................................................37
4.8.18 Install Sampler .................................................................................................38
4.8.19 Install Foam Trap.............................................................................................40
4.8.20 Plug Unused Ports ...........................................................................................41
4.8.21 Install 1.3L, 3.0L or 7.5L Fermentation Vessel Baffle.....................................42
4.8.22 Install Headplate..............................................................................................42
4.8.23 Install Vessel ....................................................................................................43
4.8.24 Install Motor Assembly.....................................................................................43
4.8.25 Make All Connections ......................................................................................43
4.9 MAIN POWER SWITCH ............................................................................................... 44
4.10 OPTIONAL BIOCOMMAND SOFTWARE ....................................................................... 45
5SPECIFICATIONS........................................................................................................47
5.1 CERTIFICATIONS........................................................................................................ 48
6OPERATING CONTROLS..........................................................................................50
6.1 TOUCHSCREEN .......................................................................................................... 50
6.2 DISPLAY SCREENS..................................................................................................... 50
6.2.1 Touchscreen Calibration..................................................................................50
6.2.2 Start-Up Screen................................................................................................51
6.2.3 Summary Screen...............................................................................................51
6.2.4 Keypads............................................................................................................54
6.2.5 Gauge Screens..................................................................................................55
6.2.6 Selecting Loop Control Modes.........................................................................56
6.2.7 Entering Loop Setpoints...................................................................................56
6.2.8 Modifying Setpoints..........................................................................................57
6.2.9 Calibration Screen ...........................................................................................58
6.2.10 Cascade Screen ................................................................................................58
6.2.11 Pump Screen.....................................................................................................59
6.2.12 Setup Screen.....................................................................................................59
7PROBE PREPARATION & CALIBRATION............................................................62
7.1 PHPROBE INSPECTION .............................................................................................. 62
7.2 PHPROBE CALIBRATION ........................................................................................... 62
7.2.1 pH Probe Installation.......................................................................................63
7.2.2 pH Probe Maintenance & Storage...................................................................65

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BioFlo/CelliGen 115 •M1369-0050 User’s Guide
7.3 DISSOLVED OXYGEN (DO) PROBE PREPARATION ..................................................... 66
7.3.1 Inspecting the DO Probe..................................................................................66
7.3.2 DO Probe Preparation.....................................................................................66
7.3.3 DO Probe Installation......................................................................................67
7.3.4 DO Probe Polarization ....................................................................................68
7.3.5 DO Probe Calibration: Setting Zero...............................................................68
7.3.6 DO Probe Calibration: Setting Span ..............................................................70
7.4 LEVEL PROBE CALIBRATION ..................................................................................... 70
7.5 ABOUT PUMP CALIBRATION...................................................................................... 71
8VESSEL STERILIZATION..........................................................................................72
8.1 INITIAL PREPARATION FOR AUTOCLAVING................................................................ 73
8.2 AUTOCLAVING THE VESSEL ...................................................................................... 74
8.2.1 Sterilization Time and Temperature.................................................................75
9REINSTALLING THE VESSEL ASSEMBLY ..........................................................76
9.1 REINSTALL THE VESSEL ASSEMBLY .......................................................................... 76
9.2 LOAD PUMP TUBING ................................................................................................. 76
9.3 CONFIRM PHCALIBRATION....................................................................................... 78
9.4 INSTALL LIQUID ADDITION SYSTEMS........................................................................ 78
9.4.1 Addition Tubing Size ........................................................................................79
9.5 RECONNECT GASES ................................................................................................... 80
9.6 INSTALL TEMPERATURE (RTD) PROBE ..................................................................... 80
10 CASCADE CONTROL .............................................................................................81
10.1 CREATING A CASCADE .............................................................................................. 82
11 ABOUT PUMPS.........................................................................................................84
11.1 PUMP ASSIGNMENT ................................................................................................... 84
11.2 PUMP SETPOINT......................................................................................................... 85
11.3 PUMP CONTROL MODE.............................................................................................. 87
11.4 PUMP FLOW RATE &CALIBRATION METHODS ......................................................... 87
11.5 PUMP PERIOD ............................................................................................................ 88
11.6 USING LEVEL PROBES TO PROGRAM FEED PUMPS .................................................... 89
11.6.1 Setting a Feed Pump to Add Liquid .................................................................89
11.6.2 Setting a Feed Pump to Harvest.......................................................................90
11.6.3 Level Control Off..............................................................................................90
11.6.4 Pump Calibration.............................................................................................90
12 USING THE SETUP SCREEN.................................................................................92
12.1 CONTROLLER SETUP ................................................................................................. 92
12.1.1 Gas Control......................................................................................................94
12.2 SYSTEM SETTINGS..................................................................................................... 97
12.2.1 Resetting Date/Time.........................................................................................97
12.2.2 Updating Software............................................................................................97
12.3 HARDWARE SETUP .................................................................................................... 98
12.3.1 Identifying Utility Station(s) Added................................................................100

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New Brunswick Scientific User’s Guide
12.3.2 Removing a Utility Station .............................................................................100
13 PERFORMING A RUN...........................................................................................101
13.1 SET UP FOAM CONTROL.......................................................................................... 101
13.2 PREPARING FOR A FERMENTATION RUN .................................................................. 101
13.3 INOCULATION.......................................................................................................... 102
13.4 START BIOCOMMAND (IF PRESENT) ........................................................................ 103
13.5 SAMPLING PROCEDURE ........................................................................................... 103
13.6 FERMENTATION PHASES.......................................................................................... 103
13.6.1 Lag Phase.......................................................................................................103
13.6.2 Exponential Growth Phase.............................................................................104
13.6.3 Steady State Phase .........................................................................................104
13.6.4 Decline Phase.................................................................................................104
13.7 BATCH OPERATION ................................................................................................. 104
13.8 FED BATCH OPERATION .......................................................................................... 104
13.9 CONTINUOUS OPERATION ....................................................................................... 105
13.10 ANAEROBIC AND MICROAEROPHILIC CULTURE .................................................. 105
13.11 HARVESTING PROCEDURE ................................................................................... 106
13.12 SHUTDOWN PROCEDURE ..................................................................................... 106
14 ESSENTIAL OPERATING TIPS ..........................................................................108
14.1 PRECAUTIONS FOR GLASS VESSEL ASSEMBLY ........................................................ 108
14.2 EXHAUST CONDENSER &EXHAUST FILTERS........................................................... 108
14.3 INSTALL A DOUBLE FILTER SYSTEM ....................................................................... 109
15 CLEANING ..............................................................................................................110
15.1 CLEANING THE VESSEL ........................................................................................... 110
15.1.1 List of Wetted Parts........................................................................................110
15.2 CLEANING THE CABINET ......................................................................................... 110
16 MAINTENANCE.....................................................................................................111
16.1 PHPROBE MAINTENANCE AND STORAGE................................................................ 111
16.2 DO PROBE MAINTENANCE AND STORAGE .............................................................. 111
16.3 VESSEL &TUBING................................................................................................... 112
16.4 PERIODIC INSPECTION ............................................................................................. 112
16.5 AGITATOR BEARING HOUSING ................................................................................ 112
16.5.1 Motor Assembly Replacement........................................................................112
16.6 REPLACEMENT PARTS ............................................................................................. 113
17 SERVICE..................................................................................................................114
17.1 TROUBLESHOOTING................................................................................................. 114
18 DRAWINGS .............................................................................................................116
18.1 LIST OF DRAWINGS ................................................................................................. 116
18.2 LIST OF TABLES....................................................................................................... 117
19 APPENDIX A: SOME GENERAL CONCEPTS.................................................118

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BioFlo/CelliGen 115 •M1369-0050 User’s Guide
19.1 WHAT IS A CONTROLLER?....................................................................................... 118
19.2 WHAT IS A CONTROL LOOP? ................................................................................... 118
19.3 WHAT IS PROBE CALIBRATION? .............................................................................. 119
19.4 WHAT ARE P-I-D CONSTANTS?............................................................................... 119
19.5 WHAT IS P-I-D TUNING?......................................................................................... 120
19.6 WHAT DO THE CONSTANTS MEAN? ........................................................................ 121
20 APPENDIX B: OTR................................................................................................122
20.1 DETERMINING AN OXYGEN TRANSFER RATE .......................................................... 122
20.1.1 OTR Calculations...........................................................................................122
20.2 SOME FACTORS THAT AFFECT OTR AND HORSEPOWER......................................... 123
21 APPENDIX C: FERMENTATION TECHNIQUES...........................................125
21.1 MEDIA FORMULATION ............................................................................................ 125
21.2 ANTIFOAM FORMULATION ...................................................................................... 126
21.3 TUBING SIZE............................................................................................................ 126
21.4 ACID &BASE .......................................................................................................... 127
21.5 GLUCOSE FEED........................................................................................................ 127
21.6 RECOMMENDED PROCESS CONTROL SETTINGS ....................................................... 128
21.7 TYPICAL FERMENTATION RUN ................................................................................ 128
21.7.1 Vessel Preparation Before Autoclaving.........................................................128
21.7.2 Vessel Sterilization.........................................................................................130
21.7.3 Post-Sterilization Vessel Set-Up.....................................................................131
21.7.4 Vessel Operation ............................................................................................132
21.7.5 Vessel Shutdown & Cleaning.........................................................................132
22 APPENDIX D: CORROSION RESISTANCE.....................................................136
23 APPENDIX E: GENERAL CHARACTERISTICS OF EPR.............................137
23.1 IDENTIFYING EPR ................................................................................................... 137
23.2 GENERAL CHARACTERISTICS .................................................................................. 137
24 INDEX.......................................................................................................................138

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New Brunswick Scientific User’s Guide
1
1 W WA
AR RN NI IN NG GS S &
&
C
CA
AU UT TI IO ON NS S
The following section is a recap of all WARNING and CAUTION messages contained in
this manual. This information is essential to the safe operation of your BioFlo/CelliGen
115. Please take a moment to acquaint yourself with the content of each message.
Page numbers are provided so you can review the message and its application within its
overall context.
1.1 WARNINGS
WARNING Page
High voltage. Always make sure this equipment is properly grounded. iv, 15
This product is not designed to contain gases within the range between
their LEL & their UEL. iv
Never block the exhaust to pressurize the vessel. 7, 37, 44
Do not use this equipment in a hazardous atmosphere or with hazardous
materials for which the equipment was not designed. 14, 16
NEVER PRESSURIZE A GLASS VESSEL!
•Always use eye protection, and exercise caution in the vicinity of glass. If the
vessel exhaust becomes blocked, pressure can build up, possible shattering
the vessel and endangering personnel.
•As soon as you open the airflow valve(s), verify by feel that air is flowing
freely from the exhaust. If not, immediately close the valve(s) or turn off the
air/gas supplies.
•Never intentionally block the exhaust to raise vessel pressure.
•Use the minimum air/gas pressure that will provide adequate airflow for the
application. Never exceed the maximum air pressure of 10 psi. This
maximum pressure is necessary only to obtain the highest gas flow rates.
18
NEVER cut any portion of the heat blanket. NEVER fold the heat blanket or
place any weight upon it. For storage, always lay the heat blanket flat. 20, 25
During autoclaving, the vessel exhaust filter must be vented to avoid explosion. 72
Use protective gloves when handling hot components. 72
Be sure to let the vessel cool…before reconnecting the water line. 76
Be careful not to pinch your fingers in the pump head levers. 78
Always turn your BioFlo/CelliGen 115 off and disconnect the power cord
before performing maintenance. 111, 114
NO ONE BUT A PROFESSIONAL SERVICE PERSON should touch electric or
electronic parts or assemblies in the electrical cabinet. 112

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BioFlo/CelliGen 115 yM1369-0050 User’s Guide
1.2 CAUTIONS
CAUTION Page
This equipment must be operated as described in this manual. If
operational guidelines are not followed, equipment damage and personal injury
can occur. … Do not use this equipment in a hazardous atmosphere or
with hazardous materials for which the equipment was not designed.
iv
Before making electrical connections, verify that the supply voltage matches
the voltage and the power requirements marked on the electrical specification
plate …and the control schematics supplied with the unit.
12, 15
Make sure all utility connections have been securely made before
connecting to WATER-IN and before turning on the main water supply. 15
Before connecting or disconnecting the water hoses to/from the vessel
and/or cabinet at any time, make sure the main water supply is closed. 16, 72,
102
To protect the integrity of your glass vessel and to avoid damage,
familiarize yourself with these cautions… 18
…do not fill the water jacket or operate the vessel until you have cut the
cable ties and released the stir bar. 26
Finger tighten only any adapter that has a white Teflon ferrule (tapered,
cone-shaped insert under the Teflon washer). The ferrule can deform under too
much pressure.
31
Make sure that the thermowell does not touch the cooling coil. 32
Do not install the pH port adaptor in the headplate before inserting the
probe... 34, 64
Do not install the dO2 port adaptor in the headplate before inserting the
probe… 35, 67
To avoid vessel stress cracks, especially during autoclaving, make vessel
clamping screws finger tight. 43
Before turning on the main power switch, make sure that: (1) the input water
hose is connected, the drain line is connected and the water supply is turned on;
(2) the vessel is in place and the quick-connect water lines are connected to the
vessel’s heat exchanger; (3) the power cord is properly connected to the control
cabinet and plugged into a suitable power outlet.
45
Be sure to wear protective gloves when installing a glass electrode. 63
We recommend that you avoid the use of hydrochloric acid (HCl)
with the BioFlo/CelliGen 115 for pH control or any other purpose, because HCl
corrodes stainless steel. Over time, it will severely damage the headplate, a
costly component to replace, and other stainless steel components.
Phosphoric and sulfuric (10% maximum concentration) acids are acceptable and
are commonly used for pH control.
65
Never let a pH probe rest on its tip, and never leave a pH probe in DI water. 65, 111
During sterilization: the bearing housing cap must be installed on the
fermentor bearing housing, to keep steam from damaging the internal
bearings.
72
Never autoclave PVC tubing (clear with white braiding). 72
...continued...

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New Brunswick Scientific User’s Guide
CAUTION Page
During autoclaving, the vessel must be vented at all times. Release the
autoclave pressure only when the temperature has dropped below 90º C. Use
slow exhaust.
75
Proper pH control is critically dependent on tubing size, which should be as
small as possible. 79
Never clean the vessel or its components or the control cabinet with abrasive
chemicals or materials. 110
Never let a DO probe rest on its tip. 111

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BioFlo/CelliGen 115 yM1369-0050 User’s Guide
2
2 I IN
NS SP PE EC CT TI IO ON N &
&
U
UN
NP PA AC CK KI IN NG G O OF F
E
EQ
QU UI IP PM ME EN NT T
2.1 Inspection of Box(es)
When you have received your order from New Brunswick Scientific, carefully inspect
all parts of the shipment for damage that may have occurred during shipping. Report
any damage immediately to the carrier and to your local NBS Sales Order Department.
2.2 Packing List Verification
Verify against your NBS packing list that you have received the correct materials.
Report any missing parts to your local NBS Sales Order Department.
2.3 Basic Components
You should have at least the following components, which will be described in greater
detail later in this manual:
•Control Cabinet with Touchscreen* •Bearing Housing
•Vessel •Filters & connectors
•Thermowell & RTD •Inoculation/Addition System
•Baffles (for fermentation only) •Sampling System
•Impellers •Harvesting System
•Probe Kits (i.e., pH, DO, Foam, Level) •Sparging System
•Motor
*While you may have multiple units, each with its own components, you will have
ordered only one touchscreen.
#NOTE:
The assembled Control Cabinet/Touchscreen assembly is called a Control
Station. For purposes of clarity in this manual, however, the control
cabinet (which houses the controller) and the touchscreen will be referred
to separately by their component names.

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New Brunswick Scientific User’s Guide
3
3 I IN
NT TR RO OD DU UC CT TI IO ON N &
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OV
VE ER RV VI IE EW W
3.1 System
BioFlo/CelliGen 115 is a versatile fermentor/bioreactor that provides a fully equipped
system in one compact package. It can be employed for batch, fed batch or continuous
culture with process control for pH, dissolved oxygen (DO), agitation, temperature,
pump feed, antifoam and foam/level.
Systems can be configured as either control stations or utility stations. Each individual
stand-alone system is a control station. One control station can run up to two
additional utility stations, which are dependent on the control station.
3.2 Vessels
One of the most versatile features of the BioFlo/CelliGen 115 is the wide variety of
glass vessels available. There are two types of vessels, non-jacketed (heat-blanketed)
and water-jacketed. Each type of vessel is available in four sizes: 1.3 liters, 3.0 liters,
7.5 liters and 14.0 liters. Ports in the headplate are provided for, but not limited to, the
following purposes: inoculation; base and acid addition; a thermowell for a resistance
temperature detector (RTD); a foam probe; a sparger; a harvest tube; a sampling tube; an
exhaust condenser; and dissolved oxygen (DO) and pH electrodes. The drive bearing
housing is also located on the headplate (see Figures 3 & 8).
3.3 Agitation System
A removable agitation motor located on top of the bearing housing on the headplate is
connected to the agitation shaft with a direct drive coupling or a magnetic coupling.
It can be easily disconnected before autoclaving the vessel and easily replaced after
sterilization. The motor will provide a speed range from 50 to 1200 RPM for
fermentation with direct drive, from 25 to 400 RPM for cell culture with direct drive,
or from 25 to 200 RPM for cell culture with magnetic drive. The process control
software ensures agitation speed control throughout the speed range.
It is possible to cascade Dissolved Oxygen (DO) to Agitation (AGIT) so the agitation
speed will vary between the user-specified minimum and maximum setpoints in order
to maintain the set percentage of DO. (See Section 10 for further information on
setting up cascades.)
Default P & I (proportional & integral) values are preset at the factory. We strongly
recommend that you maintain the factory-set parameters.

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BioFlo/CelliGen 115 yM1369-0050 User’s Guide
3.4 Temperature Control
The culture temperature setpoint may be selected within the range from 20°C above
coolant temperature to 70°C for 1.3- to 7.5-liter vessels, and from 20°C above coolant
temperature to 65°C for 14.0-liter vessels. It is controlled by the process control
software which then sends information to either a heater blanket and cooling coil or to
a water jacket. The media temperature is sensed by a Resistance Temperature
Detector (RTD) submerged in the thermowell.
Default P & I (proportional & integral) values are preset at the factory. We strongly
recommend that you maintain the factory-set parameters.
3.5 Aeration
Up to four gases, including air, nitrogen, carbon dioxide and oxygen, can be
introduced into the media through the ring sparger or optional microsparger. The flow
rate is controlled manually by one, two, three or four rotameter(s) or automatically by
thermal mass flow controller (TMFC), according to the definition of your system. The
TMFC is regulated automatically according to values set via the control station
touchscreen.
The gas mix can either be controlled manually by adjusting the flow of gases through
their rotameters or automatically if 4-gas mixing was purchased as an option. (For
further information on cascading, see Section 10.) 4-gas mixing allows the system to
automatically calculate the gas mix in response to culture needs.
Default P & I (proportional & integral) values are preset at the factory. We strongly
recommend that you maintain the factory-set parameters.
3.6 pH Control
pH is controlled in the range of 2.00-14.00. The pH is sensed by a gel-filled pH probe
(see Figures 11 & 28). Control is maintained by a P & I (proportional & integral)
controller which operates peristaltic pumps, assigned to perform acid or base addition,
or which controls the use of gas(es) for this purpose. The user can also select a
deadband value to control pH within the user-assigned range: no acid or base will be
added when the pH value falls within the deadband tolerance above or below the
setpoint.
Default P & I (proportional & integral) values are preset at the factory. We strongly
recommend that you maintain the factory-set parameters.

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New Brunswick Scientific User’s Guide
3.7 DO Control
Dissolved oxygen (DO) is controlled in the range of 0-200%. It is sensed by the DO
electrode and control is maintained by the P & I controller by changing the speed of
agitation, the thermal mass flow controller-regulated flow rate (if your system is so
equipped), and/or the percentage of oxygen in aeration.
Default P & I (proportional & integral) values are preset at the factory. We strongly
recommend that you maintain the factory-set parameters.
The DO probe is a polarographic probe. Be sure to inspect the DO probe before every
run, changing the electrolyte solution and membrane as needed.
3.8 Foam/Level Control
Foam can be monitored during batch fermentation by a foam/level probe located in the
headplate. The controller operates the antifoam-assigned pump that adds chemical
defoamer into the vessel as needed. The internal level can also be controlled by using
this feature. Pumps can be triggered to turn on or off in response to the presence or
absence of liquid.
3.9 Exhaust System
The exhaust gases pass into the exhaust condenser (see Figures 13 and 14) where
moisture is removed, then returned to the vessel. The remaining gases then pass
through a 0.2 μm exhaust filter. Be sure to inspect filters before every run, replacing
them as needed.
WARNING!
NEVER block the exhaust to pressurize the vessel!
3.10 Recommended Accessories & Supplies
Before you begin to assemble your BioFlo/CelliGen 115, it would be prudent to verify
that you have all of the following accessories and supplies readily at hand:

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BioFlo/CelliGen 115 yM1369-0050 User’s Guide
•An autoclave •An inoculation syringe
•Rubber gloves •Media
•Silicone tubing •Antifoam agent
•A tie gun •Aluminum foil
•Plastic ties (multiple colors can be helpful) •Rubber bands
•Plastic tubing connectors •pH 4 buffer
•Addition bottles •pH 7 buffer
•A liquid trap •Silicone O-ring lubricant (for
•Polysulfone quick-connects fermentation only)
User’s kits and start-up kits are available from NBS with many of the commonly
required items (including a selection of tubing, clamps, filters, connectors and addition
vessels). Speak to your NBS sales representative for more information.
3.11 Supervisory Software
In addition to the built-in software that you interface with through the touchscreen,
your BioFlo/CelliGen 115 system can be remotely controlled from a PC via NBS
BioCommand optional supervisory software (see Section 4.10). Consult your NBS
representative for details; be sure to ask for ModBus protocol.