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New Brunswick Scientific

User’s Guide

Glucose feeding can be set to respond to other sensor cues (such as DO level, the pH
reading, the turbidity measurement, the glucose measurement, etc.). The pumping
profile to be used must generally be determined through experimental experience.

23.6

Recommended Process Control Settings

Question: What are the recommended process control settings (i.e., temperature,
pH, agitation speed, DO & gas sparge rate)?

Answer:

For

E. coli,

temperature is usually set to 32° - 35°C and pH is set at 7.0 - 7.2.

For yeast the values are 30°C and a pH value of 5.0. Agitation speed is usually set to a
minimum of 200 - 300 rpm with a maximum value of 1000 rpm. Dissolved oxygen
(or DO) level is usually 30%. The gas sparge rate is generally 0.5 to 1.0 VVM.

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Summary of Contents for BioFlo 415

Page 1: ...P Fermentor MANUAL NO M1360 0050 Revision D December 15 2008 NEW BRUNSWICK SCIENTIFIC BOX 4005 44 TALMADGE ROAD EDISON NJ 08818 4005 Telephone 1 732 287 1200 1 800 631 5417 Fax 732 287 4222 bioinfo nbsc com Web www nbsc com ...

Page 2: ...ad Shanghai 200235 P R China Tel 86 21 6484 5955 or 5966 Fax 86 21 6484 5933 E mail nbschc online sh cn THE NETHERLANDS New Brunswick Scientific BV Kerkenbos 1101 6546 BC Nijmegen P O Box 6826 6503 GH Nijmegen Nederland Tel 31 0 24 3717 600 Fax 31 0 24 3717 640 E mail sales nbsbv nl FRANCE New Brunswick Scientific SARL 12 14 Rond Point des Champs Elysées 75008 Paris France Tel 33 0 1 5353 1511 Fax...

Page 3: ...ted 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 ...

Page 4: ...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 ...

Page 5: ... 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...

Page 6: ..._______________________ SERIAL NUMBER ________________________________ The above information can be found on the electrical specification plate Purchased with the following installed options __________________________________________________ __________________________________________________ __________________________________________________ __________________________________________________ _____...

Page 7: ... 11 RECOMMENDED ACCESSORIES SUPPLIES 7 3 12 SUPERVISORY SOFTWARE 7 4 INSTALLATION 8 4 1 PHYSICAL LOCATION 8 4 2 ENVIRONMENT 8 4 3 INSTALLING THE CONTROL CABINET 8 4 4 INSTALLING THE TOUCHSCREEN 8 4 5 UTILITIES 10 4 5 1 Electrical Requirements 11 4 5 2 Water and Drain Connections 12 4 5 3 Gas Connections 12 4 5 4 Exhaust Connection 12 4 6 VESSEL ASSEMBLY 13 4 6 1 Sparge Exhaust Assemblies 15 4 6 2 ...

Page 8: ...BRATION 42 7 1 PH PROBE INSPECTION 42 7 2 PH PROBE CALIBRATION 42 7 2 1 pH Probe Installation 44 7 2 2 pH Probe Maintenance Storage 45 7 3 DISSOLVED OXYGEN DO PROBE PREPARATION 45 7 3 1 Inspecting the DO Probe 45 7 3 2 DO Probe Preparation 46 7 3 3 DO Probe Installation 46 7 3 4 DO Probe Polarization 47 7 3 5 DO Probe Calibration Setting Zero 47 7 3 6 DO Probe Calibration Setting Span 48 8 AUTOMAT...

Page 9: ...DE 67 11 2 CONTROLLING DO BY CASCADE 69 12 PLOTTING TRENDS 72 12 1 CREATING A TREND GRAPH 72 12 1 1 Using the Export Button 74 12 1 2 Using the Zoom Button 75 12 1 3 Using the Read Line 75 13 ABOUT PUMPS 77 13 1 PUMP SETPOINT 77 13 2 PUMP CONTROL MODE 78 13 3 PUMP FLOW RATE CALIBRATION METHODS 79 13 4 PUMP PERIOD 80 13 5 INSTALLING AN EXTERNAL VARIABLE SPEED PUMP 80 14 ABOUT ALARMS 82 14 1 ABS AND...

Page 10: ...se 105 16 8 BATCH OPERATION 105 16 9 FED BATCH OPERATION 105 16 10 CHEMOSTAT OPERATION 106 16 11 SHUTDOWN PROCEDURE 107 17 CLEANING 108 17 1 CLEANING THE VESSEL 108 17 1 1 List of Wetted Parts 108 17 2 CLEANING THE CABINET 109 18 MAINTENANCE 110 18 1 PH PROBE MAINTENANCE AND STORAGE 110 18 2 DO PROBE MAINTENANCE AND STORAGE 110 18 3 VESSEL TUBING 111 18 4 PERIODIC INSPECTION 111 18 5 FUSE REPLACEM...

Page 11: ...ER RATE 126 22 1 1 OTR Calculations 126 22 2 SOME FACTORS THAT AFFECT OTR AND HORSEPOWER 127 23 APPENDIX C FERMENTATION TECHNIQUES 128 23 1 MEDIA FORMULATION 128 23 2 ANTIFOAM FORMULATION 129 23 3 TUBING SIZE 129 23 4 ACID BASE 130 23 5 GLUCOSE FEED 130 23 6 RECOMMENDED PROCESS CONTROL SETTINGS 131 24 APPENDIX D CORROSION RESISTANCE TABLES 132 25 APPENDIX E GENERAL CHARACTERISTICS OF EPR 147 25 1 ...

Page 12: ......

Page 13: ...rforming a sterilization cycle 49 Be sure to wear protective gloves when you move the sparge valve from sterilization to sparge position 54 Never turn the heater off during a run or the exhaust filter may clog 66 Be careful not to pinch your fingers in the pump head levers 100 Always turn your BioFlo 415 off and disconnect the power cord before performing maintenance 110 113 1 2 CAUTIONS CAUTION P...

Page 14: ...ric and sulfuric 10 maximum concentration acids are acceptable and are commonly used for pH control 45 Never let a pH probe rest on its tip and never leave a pH probe in DI water 45 110 There must be liquid in the vessel for sterilization Never begin the sterilization process if the vessel is empty 49 If Sterilization Temperature does not reach 121 5ºC make sure the water supply pressure is not to...

Page 15: ...r 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 Filters Touchscreen Motor Vessel components installed Hoses Tubing Sampling Harvesting Connections Probe Kits pH DO Foam...

Page 16: ...ance 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 3a 8 9 and 10 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 magnetic drive coupli...

Page 17: ...ng to values set via the control cabinet touchscreen If you wish to use a rotameter there is an option for no thermal mass flow controller The percentage of oxygen blended with the sparge air can be controlled manually by the user or automatically through the controller by applying the O2 enrichment function For further information on cascading see Section 11 3 6 pH Control pH is controlled in the...

Page 18: ...Foam can be controlled during batch fermentation by a level probe located in the headplate The controller operates the antifoam assigned pump that adds chemical defoamer into the vessel as needed 3 9 Exhaust System The exhaust gases pass into the exhaust condenser see Figure 5 where moisture is removed any residual moisture is heated past the dewpoint via the heating pad allowing it to pass throug...

Page 19: ...ng lubricant Addition bottles An inoculation syringe A liquid trap A spare parts kit is available from NBS part number M1360 9969 see Section 19 2 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 12 Supervisory Software In addition to the built in software that you i...

Page 20: ...C Relative humidity up to 80 non condensing 4 3 Installing the Control Cabinet Position the BioFlo 415 control cabinet on a firm level surface in an area where utilities are readily available Level the horizontal surface of the base with four leveling glides if necessary Connect the power cord to the rear of the control cabinet At a later time once the unit is completely assembled and all connecti...

Page 21: ...hes the voltage and the power requirements marked on the electrical specification plate located on the rear panel of the cabinet and the control schematics supplied with the unit The unit is shipped with the gas inlets plugged Be sure to remove each metal plug before you insert tubing IF ANY GAS INLET WILL NOT BE USED KEEP IT CLOSED OFF with the metal plug provided ...

Page 22: ... power and an open drain All service connections are located on the lefthand side of the cabinet see Figure 1a Using standard plant practices and respecting all applicable codes connect services to the appropriate connections as recapped in Table 1 on the following page and explained in greater detail in Sections 4 5 1 4 5 3 TOUCHSCREEN Rear View Attach the monitor to the Control Cabinet mounting ...

Page 23: ...IG Water Supply Quick connect 10 PSIG Water Return Quick connect Max 5 PSIG Power 208 230 15 Amp 50 60 Hz NOTE When you connect gas and water we recommend that you install pressure regulators to control the pressures as recommended in the utility chart Table 1 4 5 1 Electrical Requirements 208 230 Volts 50 60 Hertz 15 Amp CAUTION Before making electrical connections verify that the supply voltage ...

Page 24: ...t was not designed All gases supplied should be medical grade Never leave a gas inlet open if no tubing will be connected keep the inlet plugged Not for use with gases in the range between their LEL and their UEL All gases should be regulated using a two stage regulator The scale of the regulator gauge for gases going into the fermentor should be such that one can regulate pressure between 0 30 PS...

Page 25: ...ction properly each system needs to be set up as explained in this section Failure to follow these instructions will result in the system s inability to function and will void the warranty NOTE When installing the equipment it is best to choose a location that has access to the necessary utilities and a drain for water return and exhaust As you set up the BioFlo 415 the control cabinet must be pos...

Page 26: ... SPARE PORT SINGLE SEPTUM 12mm PORT HARVEST SAMPLE EXHAUST THERMOWELL LEVEL 12mm PORT HARVEST SAMPLE VESSEL LIGHT 12mm SPARE PORT SINGLE SEPTUM PG 13 5 PORT DO pH PG 13 5 PORT DO pH 19mm SPARE PORT MULTIPLE SEPTUM LEVEL 12mm SPARE PORT SINGLE SEPTUM 4 C SINK HOLES FOR BAFFLE PLUGS SPARGER CAUTION Never use a wrench on headplate ports only finger tighten the adapters If a leak should be detected at...

Page 27: ... LEVEL 12mm PORT HARVEST SAMPLE 12mm SPARE PORT SINGLE SEPTUM EXHAUST CAUTION Never use a wrench on headplate ports only finger tighten the adapters If a leak should be detected at a port replace the O ring 4 6 1 Sparge Exhaust Assemblies Sparge Assembly This assembly see Figure 4 consists of the sparge valve sparge line inlet filter and condensate return line The assembly should be installed in o...

Page 28: ...se lines that were removed prior to assembly 2 Make sure the sample septum tri clamp cap is in place on the harvest sample tube assembly 3 Make sure the exhaust condenser s heater pad is plugged in see Figure 1a INLET FILTER install filter into SIP housing CONDENSATE RETURN connect cable to FY2A on side of cabinet SPARGE VALVE Be sure to read WARNING in Section 8 2 3 Pull hold the ring out Lift th...

Page 29: ...port 1 Apply a small amount of vacuum grease to the external threads of the adapter plug then insert the plug into the available 19mm port on the headplate See Figures 3a 3b for recommended locations 2 Tighten the plug until it bottoms out in the port 3 Insert the probe into the adapter plug until the threads on the probe engage into the top of the plug 4 Tighten the probe into the adapter until t...

Page 30: ...ns are intact Set the Temp setpoint see Section 10 4 1 10 C below process value Turn Temp to AUTO for 3 minutes see Section 10 1 to run 100 cooling The cooling water will drive out any air that was left in the water system lines and the vessel heat exchanger CAUTION On initial set up prior to operation check that all impellers are tightly fastened to the drive shaft 4 6 5 Connecting Vessel to Cabi...

Page 31: ...t at the location labeled FILTER CONDENSATE 4 Connect the Control Cabinet s LOWER JACKET WATER to the lower quick connect on the vessel jacket see Figure 7 This is used to supply water to the vessel jacket 5 Connect the Control Cabinet s UPPER JACKET WATER to the upper quick connect on the vessel jacket see Figure 7 This is used to channel water out of the jacket Figure 7 Vessel Water Inlet Outlet...

Page 32: ...l on the cabinet see Figure 1a repeated on the following page 9 Connect the vessel light cable to the VESSEL LIGHT JACK on the cabinet panel see Figure 1a 10 Connect the exhaust heater see Figure 5 to the EXH HEATER connector on the side of the cabinet 11 Confirm that the rupture disk in back of the vessel and the safety release valve in back of the vessel jacket are connected to tubing or hoses t...

Page 33: ...r inlet on the right side panel of the control cabinet then plug the cord into a suitable grounded power supply outlet The control cabinet is turned on and off with the ON OFF power switch located on the front panel in the lower right corner CAUTION As soon as the heater is connected to the Control Cabinet you must make sure there is water in the vessel jacket Do not plug the heater cable into the...

Page 34: ...3 Communications to BioCommand Plus software are via an optional RS 232 interface cable 1 Connect the 25 pin end of the RS 232 cable to the AFS Modbus port and ensure that the connection is secure 2 Hand tighten the thumbscrews 3 Refer to the BioCommand Plus user s guide for instructions on connecting the RS 232 interface cable to the supervisory host computer 4 10 Inputs Outputs for Ancillary Dev...

Page 35: ... available on the control cabinet rear panel see Figure 8 for the connection of serial ancillary devices such as scales for vessel and addition bottles You can connect a box with four serial RS232 inputs and outputs to one USB port to allow you to connect and control up to four scales or other ancillary equipment ...

Page 36: ...re Sensor Platinum RTD probe Drive Permanent magnet motor with high torque input Indication Digital display in 1 RPM increments Range 50 1000 RPM Control 1 Bearing Housing Standard single mechanical seal with magnetic drive Sensor Optical photoplastic disc 500 lines rev with quadrature output Agitation Impellers Six bladed Rushton turbine impellers provided Filter 1 2μm depth filter 0 2μm absolute...

Page 37: ... 47 lbs 21 kg 60 lbs 27 kg 80 lbs 36 kg Overall Dimensions with Touchscreen 25 in wide X 24 in deep X 34 in high 63 cm W X 61 cm D X 86 cm H External Computer Connections Port supplied for remote connection of interface computer BioCommand Plus Connections Port supplied for connection of BioCommand Plus supervisory host computer Fuses One 5A glass tube fast acting fuse Regulatory Compliance See Se...

Page 38: ...26 BioFlo 415 y M1360 0050 User s Guide All BioFlo 415 vessels have received the ASME UM stamp of approval and meet the standards of the EU Pressure Equipment Directive 97 23 EC ...

Page 39: ...on The first time you power up you will be prompted to calibrate the screen to your touch Follow the onscreen instructions to touch the target each time it appears Usually you will be prompted to touch the four corners of the screen twice in succession NOTE For optimal results be sure to stand or sit in the position from which you are most likely to work Height and angle of reach will affect calib...

Page 40: ...ou turn on the power if you have already calibrated the touchscreen see Section 6 2 1 Figure 10 Start Up Screen This screen remains in view for a few seconds then it is replaced by the SUMMARY Screen 6 2 3 Summary Screen The SUMMARY screen see Figure 11 on the following page is command central it puts as many as 32 loops at your fingertips for programming fermentation runs New Brunswick Scientific...

Page 41: ...ent output for each loop This is an automatic control function to maintain current readings within the setpoint tolerance range Control Mode Depending on the loop the control mode may be Off Auto Manual On or O2 Enrich Unit of measure This is the unit of measure used for the PV and Setpoint Cascade If any cascades have been programmed they will be displayed here continued SCROLL DOWN buttons SCROL...

Page 42: ...y and acknowledge any alarm Sterilize This screen allows you to set and monitor the parameters for the sterilization sequence Setup Use this screen to set up your PC as an auxiliary part of your BioFlo 415 system Other Buttons Description Scroll Up Press this button to scroll upwards one loop at a time Scroll Down Press this button to scroll downwards one loop at a time Vessel Light Press this but...

Page 43: ...icates valve status red CLOSED green OPEN Touch valve icon to go to STERILIZATION screen LEVEL PROBE color indicates probe status Red DRY Green WET LOOP each gauge indicates setpoint SP and process variable PV Title color indicates loop status Red OFF Green ON Blue MANUAL Touch loop gauge to go to full gauge screen see Section 6 2 5 LoopName Units The action of this loop Agitation is measured in R...

Page 44: ...ad see Figure 14a to name the loop 4 Press the appropriate option button The corresponding measurement Unit will automatically appear in this sample screen 5 Press the appropriate input device designation 6 Press the appropriate output device designation 7 Input the desired control settings 8 After making all your selections press the Õ OK button to save them Press Caps Lock to shift to CAPITAL le...

Page 45: ...he Add User Defined Loop screen that opens press the Remove button see Figure 14 4 If the loop is a pump only optional pumps have a Settings button that provides access to their Remove button see Figure 16 Figure 16 Deleting a Pump Control Loop 5 When you return to the SUMMARY screen the loop will be deleted PRESS HERE to open the Add User Defined Loop screen Press the Settings button to open the ...

Page 46: ...uaint you with the primary screens accessed from the blue buttons at the bottom of each screen 6 2 9 Calibration Screen This screen is used to calibrate the pH and the DO probes For details on probe calibration see Sections 7 2 pH probe and Error Reference source not found DO probe Figure 17 Calibration Screen These last two loops are input from the Level probes to the Level HiFoam loops 2 Press t...

Page 47: ...gs For details on setting cascades see Section Error Reference source not found Figure 18 Cascade Screen 6 2 11 Trend Screen This screen see Figure 19 allows the user to set the parameters for plotting trend graphs and to view the graphs that track the activity of up to 8 selected loops during an entire process run The data can be exported through the USB port in Excel format to a PC For details o...

Page 48: ...providing both current readings and the opportunity to change pump settings For details on using the PUMP SCREEN see Section 10 2 Figure 20 Pump Screen 6 2 13 Alarm Summary Screen This screen see Figure 21 allows the user to turn alarms on and off to read the alarm history and to acknowledge any alarm while it is active For details on using the ALARM screen see Section 14 Figure 21 Alarm Summary S...

Page 49: ...e Parameter Definition YES NO Drain Time Amount of time valves are open to prime the jacket 9 Heat A Vessel contents are heated via band heater 9 Heat B Temp Temperature at which steam is passed through filters to condensate connections 9 Sterilization Temp Temperature at which vessel is held for sterilization 9 Sterilization Time Time vessel is held at sterilization temperature 9 Cool A Time from...

Page 50: ... button the screen that opens is actually the first tab the CONTROLLER SETUP see Figure 23 screen Figure 23 Controller Setup Screen Figure 24 Recipe Manager Screen TAB The UNIT NAME is user selected Press this box then use the pop up touchpad to type in the name Use this pane to identify any additional DO options your system may have a second DO and pH option DO2 and pH2 or a second DO and Redox o...

Page 51: ...FS Modbus Use this pane to change Date and Time see Section 15 3 1 English is the default language When other choices Français Deutsch Español become available the user will select the language here Use this pane to calibrate the touchscreen see Section 15 3 Use this pane to view the Software Firmware version installed and to update Software via the USB port see Section 15 3 2 Use this screen to v...

Page 52: ... and log the current values of any parameters temp pH DO air flow pump flow rate levels and agitation that are monitored The data can also be stored plotted and afterwards transferred to other commonly available programs to be manipulated and analyzed in various ways Table 4 on the following page identifies the pin designations for this 25 pin RS232 422 connector ANALOG INPUTS OUTPUTS are easily a...

Page 53: ...ntor 7 GND Ground reference for all signals 12 IRXD 24 IRXD RS422 paired data input to fermentor 13 ITXD 25 ITXD RS422 paired data output from fermentor 21 IOS Open selects RS232 Grounded selects RS422 Unless otherwise requested the baud rate is factory selected at 9600 AFS or 19200 Modbus and the connector is configured as an RS232 port i e no jumper between pin 7 and pin 21 The factory set addre...

Page 54: ...e are filled with the same reference electrolyte for a total volume of approximately 30 mL For normal operation remove the rubber T stoppers if the probe is so equipped saving them for use during sterilization 7 2 pH Probe Calibration NOTE Calibrate the pH probe before sterilizing it in the vessel 1 If you have not already done so connect the pH probe to the pH connector on the control cabinet see...

Page 55: ...n the CURRENT VALUE reading stabilizes press the SET SPAN button 13 To ensure accuracy repeat Steps 4 11 a few times using the same two buffer solutions NOTE The pH calibration should be checked after sterilization immediately prior to inoculation Take a sample from the vessel and compare the pH value displayed on the control cabinet screen to the pH recorded by an external pH meter Any discrepanc...

Page 56: ...in the headplate 1 Apply a small amount of silicone grease or glycerol to the electrode body 2 Install the pH electrode as shown in Figure 27 Gently turn the probe as you press it into the port to ease it into the vessel without forcing Also make sure that there is no interference inside the vessel Figure 27 pH Probe Installation gel filled pH PROBE O RINGS HEADPLATE ...

Page 57: ...air bubbles in the electrode s tip to remove bubbles hold electrode upright and shake electrode gently The probe should be stored standing upright The electrode tip should be immersed in the solution of 3 molar KCl or a buffer solution between pH 4 00 and pH 7 00 If the probe is so equipped the two rubber T stoppers should be inserted CAUTION Never let a pH probe rest on its tip and never leave a ...

Page 58: ...other people have access to the unit NOTE It is recommended that you use the factory set P I values Do not attempt to change the settings unless you are very experienced with P I control 7 3 3 DO Probe Installation Install the DO probe into the vessel headplate assembly see Figures 3a 3b and Figure 28 on the following page ensuring that there is no interference inside the vessel and taking care to...

Page 59: ...han 5 minutes it will need to be re polarized To re polarize Connect the probe to the operating O2 amplifier or polarizing module Allow SIX HOURS FOR POLARIZATION prior to calibrating the probe 7 3 5 DO Probe Calibration Setting Zero NOTE The DO probe is calibrated after sterilization There are two methods to obtain zero for calibrating the DO probe Review both methods and use the one you prefer M...

Page 60: ...inutes the current value reading will stabilize 5 Press the SET ZERO edit box use the touchpad to enter 0 press the OK button then press the SET ZERO button 6 Press N2 3 OFF or if in Step 3 you manually turned the N2 loop on now manually shut off the nitrogen flow to the vessel 7 3 6 DO Probe Calibration Setting Span 1 In the AGIT GAUGE screen set the AGIT speed to 50 RPM 2 Set the AGIT mode to AU...

Page 61: ...empty CAUTION If Sterilization Temperature does not reach 121 5 C make sure the water supply pressure is not too high max 10 PSIG If the pressure is too high the water will boil off through a relief valve during the Heat A phase causing the water jacket to vapor lock with the heated regulator acting as a blocking valve If this happens lower incoming water pressure 8 1 ESSENTIAL Backflow Prevention...

Page 62: ... this connection Figure 30b NBS Backflow Prevention Kit 3 If your water pressure regulator kit was not purchased from NBS some additional pipe nipples are supplied to facilitate connection of the backflow prevention kit If additional pipe fittings are required please contact NBS or your local plumbing supply Any questions or concerns can be answered by calling New Brunswick Scientific at 1 732 287...

Page 63: ...he vessel jacket release valve lines are going to the drain 7 Confirm that the sparge inlet and exhaust outlet filters are placed in the filter housings NOTE Filters must be replaced after 8 10 sterilization cycles 8 Insert any probes the calibrated pH probe and or the DO Foam and or Level probes into the vessel Connect the probe cables to the proper jacks on the cabinet 9 Verify that mineral oil ...

Page 64: ...r the cap NOTE Be sure to move the sparger tube back into the sparge position and to open the exhaust condenser needle valve when sterilization is over The unit is now ready to be sterilized Figure 30c Moving Sparge Valve for Sterilization 8 2 1 Priming the Water Jacket To fill the water jacket 1 After the tubing and water supply have been connected make sure the RTD cable is plugged into the cont...

Page 65: ...rol loop for the desired operating temperature after sterilization Turn the Temperature control loop ON by setting the temperature control loop to AUTO control mode 2 Set the sparge gas flow rate at the desired setpoint for operation after sterilization The GasFlo control loop should be set to the AUTO control mode NOTE If you are using Gas Mixing the O2 loop must be in the O2ENRH mode 3 Ensure al...

Page 66: ...nected to the controller This valve pulses during sterilization mode to ensure sterilization of the inlet and exhaust filter NOTE The sample valve should be sterilized separately in an autoclave then aseptically connected to the unit in a clean environment 8 When the sterilization hold phase Steril Time ends the operator should reopen the needle valve on the water return line from the exhaust line...

Page 67: ...ring 8 2 4 Resetting Sterilization Parameters NOTE The sterilization parameters have been preconfigured at the factory The operator may make changes to the sterilization parameters by following the instructions below The user definable sterilization parameters are shown in Figure 31 which is a detail from the STERILIZATION screen shown in Figure 22 They are further explained in Table 5 which follo...

Page 68: ...actory set to 45 minutes Cool B Temperature Press the COOL B edit box to input the temperature at which the vessel will transition from Cool A into Cool B phase See important NOTE below Growth Temperature Press the GROWTH TEMP edit box to define the temperature at which the vessel will transition into the Growth phase NOTE Entering a Heat B temperature below 100o C will cause excessive heating tim...

Page 69: ...steam is generated in the vessel Steril Heating operation at sterilization temperature Cool A Cooling with temperature 10ºC above normal operation temperature setpoint Cool B Cooling with temperature 10ºC above normal operation temperature setpoint The factory settings for the sterilization sequence are indicated in Figure 32 which is a detail from the STERILIZATION screen Figure 32 Sterilization ...

Page 70: ... been started by pressing the Stop button in the STERILIZATION screen Upon aborting the sterlization sequence or if there is a power failure the unit will proceed to the Cool A phase and eventually return to Growth once it has cooled down Drain Phase Heat A Phase Heat B Phase Sterilization Hold Phase Cool A Phase Cool B Phase Growth Phase Process Control is indicated in black white It indicates th...

Page 71: ... putting the temperature loop in PID mode for 2 minutes Then return to the growth temperature setpoint 3 Confirm that all O rings gaskets seals filters and septa are in place lightly coated with silicone grease and in good condition 4 Make sure that the RTD TEMPERATURE PROBE is securely in place in the vessel s thermowell CAUTION Proper installation of the RTD is essential to temperature control 5...

Page 72: ...y to operate 9 2 Adding Media to the Vessel Media is usually added to the vessel via the 19mm port located on the vessel headplate 1 Most fermentation process media is added to the vessel prior to sterilization 2 Cell culture media is often composed of a variety of heat labile components If filter sterilized media is to be added to the vessel the vessel may be sterilized with phosphate buffered sa...

Page 73: ...ally associated with a different sensor The onscreen control mode choice will be the name of the loop chosen to have influence on the actuator 10 2 Pump Assignment The user has the ability to assign each pump present in the system To assign a pump 1 From any screen press the PUMPS button at the bottom to open the PUMP screen see Figure 20 repeated here for easy reference Figure 20 Pump Screen 2 Pr...

Page 74: ...pad NOTE If you change P I values you can return to the original settings at any time by pressing the Factory Default button see Figure 13 10 4 Loop Setpoints The setpoint is the value you want each loop to attain When the loop control mode is AUTO the fermentor will automatically make appropriate adjustments to maintain the value at the setpoint 10 4 1 Entering Setpoints To enter a setpoint for a...

Page 75: ...e until the DO setpoint is reached Conversely when the actual DO value drops below the setpoint the agitation speed is increased until the DO setpoint is reached This is all controlled by the cascade set up See Section 11 for details about setting cascades 4 Use the touchpad number keys to enter the desired setpoint Use the white CLEAR button at any time before Step 5 to empty the setpoint edit bo...

Page 76: ... pump s ASSIGNMENT button to open the PUMP ASSIGNMENT screen see Figure 35 repeated here for easy reference Figure 35 Pump Assignment Screen In DRY control mode when the liquid is not in contact with the probe the feed pump is turned on so that more liquid will be added when the liquid is in contact with the probe the pump is turned off 10 6 2 Setting a Feed Pump to Harvest A level probe can also ...

Page 77: ...hich should be as small as possible Consult Table 9 for guidance 10 6 4 Pump Calibration Pump flow rates are provided in Table 9 Section 13 3 However more accurate flow rates through the various lines may be established by pre calibrating the pumps using the PUMP screen This screen controls all pump parameters for the three standard fixed speed pumps supplied with each control cabinet and for any ...

Page 78: ... TMFC is installed 1 Go to the CASCADE screen and select the GasFlo loop as the Cascade From loop 2 Enable the default cascade to the exhaust heater 3 Go to the SUMMARY or SYNOPTIC screen and select the ExhstHtr loop Set it in the MANUAL control mode For all other set ups 1 Go to the SUMMARY or SYNOPTIC screen and select the ExhstHtr loop Enter a setpoint of 85 2 Set it in the MANUAL control mode ...

Page 79: ...imultaneously is used to smooth the transition from one loop to another 11 1 Creating a Cascade Figure 18a below shows the headers from the CASCADE screen set to Cascade From DO with an explanation of the settings those headers represent Figure18a Cascade Screen End Setpoint is the loop value the user defines as the maximum allowable value when the DO End Out is reached Typically this value will a...

Page 80: ...tween the present value PV the setpoint and the DO output percentage Out When the PV is greater than the setpoint the system will be generating a negative Out because the controller senses a need to decrease DO When the PV is less than the setpoint the system will be generating a positive Out because the controller senses a need to increase DO When the PV equals the setpoint the Out should be appr...

Page 81: ...able button to select YES 1 DO is the source of the cascade Agit is the loop being influenced by the cascade Start Setpoint is the value the system will set the loop setpoint to when the DO Start Out is reached This value is typically close to or at the normal operating setpoint DO Start Out is the DO output value at which the user wants the cascade to begin When this output value is reached the l...

Page 82: ...o 100 When to use 100 as DO Start Out in a Cascade Commonly after calibrating DO probes the system will have a DO present value higher than the setpoint This happens because of calibration methods and because the culture has not had enough time or has not grown to a significant enough density to start to consume the available DO This difference as described earlier will cause the DO output to plum...

Page 83: ... any cascade to loop will not go below its own Minimum Setpoint value Minimum Output corresponds to the minimum value that will produce the minimum setpoint lower outputs will not affect setpoint Regardless of DO output the setpoint of any cascade to loop will not rise above its own maximum setpoint ...

Page 84: ...selected color The graph and data are only available while the fermentor is running Data cannot be stored in the controller but can be archived remotely on an auxiliary PC via the RS232 422 Modbus interface see Section 4 9 for details or saved to a USB storage device 12 1 Creating a Trend Graph 1 From any screen press the TREND button to open the TREND screen see Figure 19 repeated below for easy ...

Page 85: ...tom of the graph Figure 39 Trend Graph TREND GRAPH BUTTONS Ö RAMP DOWN RAMP UP Touching the PV SP or OU box will open the loop gauge screen see Section 6 2 5 3 Select the first loop The program will automatically place it in the red box 4 If you wish to change the color of this loop press the new color choice here 5 Press the Display High box to enter using the touchpad the high limit for the Y ax...

Page 86: ...on to open an interactive mode where you can zoom in on a section of interest on one plot The button turns red when you touch it indicating you are in Zoom mode See Section 0 below for detailed instructions Read Line Press this button to open an interactive mode where you can move a vertical cross sectional line across the graph to aid in determining a particular reading See Section 12 1 3 below f...

Page 87: ... read line see Section 12 1 3 at the same time you must enter Zoom mode first 12 1 3 Using the Read Line The Read Line mode allows you to read PV values from the graph displayed at the top of the screen at a position you select To use the Read Line 1 Press the Read Line button at the bottom of the TREND screen It will turn red to indicate that the read line mode is active and black vertical line w...

Page 88: ...e Location 4 Press the Read Line button again to return to the regular trend graph If you wish to use the zoom mode see Section 12 1 1 and the read line at the same time you must enter Zoom mode first Press anywhere along the desired vertical axis to locate the read line ...

Page 89: ... control cabinet see Figure 42 Remember to set up any optional pumps you may have added to your system see Section 13 5 to install an external variable speed pump Figure 42 Standard Pump Array 13 1 Pump Setpoint The setpoint defines how long the pump will run with reference to the pump period see Section 13 4 To enter a setpoint for any pump 1 Open the Pump Screen Gauges for Pumps 1 3 are displaye...

Page 90: ...will run continuously When you release the button the pump will stop running To enter select a Control Mode for any pump press the appropriate button in the Control Mode pane of the PUMP screen see Figure 43 NOTE If pumps are linked to a cascade this may affect the ability to manually change setpoints and control modes 2 Press the Setpoint edit box for Pump 1 3 Use the touchpad that opens to enter...

Page 91: ... you do it with the input end of the tubing in the water reservoir prime the tubing line by pressing the pump s Prime button but allow it to run only until liquid starts to flow into the tubing DO NOT allow the liquid to run into the graduated cylinder yet 4 If you are not using a scale skip to Step 5 If you are using a scale place the graduated cylinder with the tubing on the scale and press Zero...

Page 92: ... 3 5 seconds then cycle back on again NOTE Running at a very low percentage renders the totalizer s results inaccurate We recommend the use of smaller tubing to avoid choosing a very low percentage for the pump setpoint 13 5 Installing an External Variable Speed Pump CAUTION When selecting an external pump to operate with your system please consult with your local sales representative to be sure t...

Page 93: ...ve output at the top be sure to use the same number 1 2 or 3 as you used for the green wire 5 If and only if you are using a 0 5V connection instead of 4 20mA connect the end of the green wire return to one of the four 4 5 6 or 7 negative outputs at the bottom and connect the end of the black wire 0 5V input to the matching positive outputs at the top 6 Set up the pump control loop using a loop fo...

Page 94: ...red when the control loop s Process Variable falls below or rises above the control band that you specify around the loop s setpoint e g a tolerance of 10 rpm above or 5 rpm below the Agitation setpoint 14 2 Setting Alarms To set alarms 1 Press the ALARMS button to open the ALARM screen see Figure 21 repeated here for easy reference and Table 10 which explains the features of this screen Figure 21...

Page 95: ... loops setpoints DEVHigh This column indicates any tolerance you have set above the control loops setpoints DEVEnable DEVAudible This column indicates whether the Deviation alarm limits have been enabled Active or not InActive for visible DEVEnable and or audible DEVAudible alarms Acknowledge All button Press this button to acknowledge and stop all alarms Current Alarms button Press this button to...

Page 96: ...rm e g Unit1 Deviation Low Error The Visible alarm will remain onscreen until the alarm condition is rectified If the Audible alarm is also enabled beeping will occur until the alarm is acknowledged There are three ways to acknowledge alarms 1 one alarm at a time 2 all alarms for one control loop at a time and 3 all alarms for all control loops at a time for the rare occasion such a condition shou...

Page 97: ...LARM screen 2 Press the Acknowledge All button All alarms will be deleted from this screen 3 Press the Scroll Back button to return to the ALARM screen CAUTION Acknowledging alarms is NOT a replacement for correcting the condition that triggered the alarm Diagnose the cause of the alarm condition and rectify the situation to ensure proper operation of your BioFlo 415 14 4 Alarms History Each time ...

Page 98: ...pName box in the main ALARM screen 2 In the control loop s Alarms Screen that opens press the Alarm History button 3 Press the Purge button to erase all records NOTE You cannot delete one record at a time you can only purge all records simultaneously 4 Press the Scroll Back button to return to the ALARM screen A sample screen may look like this 1 Temp 1 28 2007 9 21 15 AM Deviation Low Error Activ...

Page 99: ...the Hardware Setup see Section 15 4 In addition if you need to contact NBS Customer Service about your BioFlo 415 you may wish to access this screen to check in the Hardware Setup pane the status of installed modules and the firmware version which you also see briefly in the START UP screen Figure 23 Controller Setup Screen 15 1 Controller Setup When you open the SETUP screen normally the Controll...

Page 100: ...s loop gauge screens Finally you can also use this screen to change your auxiliary pH DO module selection by pressing the other option button The Save Changes button saves your new selections and reconfigures all control loops accordingly Although you can save each change one at a time in this screen by pressing it you can also wait until all changes have been selected If you leave this screen how...

Page 101: ...Gas the process loops are labeled Air 1 O2 2 and CO2 4 The loops numbers 1 2 4 correspond to the gas connections on the cabinet see Figure 1a There is also a GasFlo loop when one TMFC is present settings in this loop s gauge screen turn the TMFC on and off and control the gas flow rate 15 1 2 Gas Control with 2 TMFCs If your system is equipped with two TMFCs the only Operating Mode available in th...

Page 102: ...de see Figure 53 below Figure 53 Controller Setup Screen 3 TMFCs The gas process loops you will find in the SUMMARY screen are labeled AirFlo 1 O2Flo 2 and CO2Flo 3 The loops numbers 1 3 correspond to the gas connections on the cabinet see Figure 1a With 2 TMFCs you can only select O2 Enrich as the Operating Mode 3Gas mode is grayed out inaccessible With 3 TMFCs you can select O2 Enrich or 3Gas as...

Page 103: ...lso a third loop called Gs3Flo 3 for the addition of a gas of your choosing The loops numbers 1 4 correspond to the gas connections on the cabinet see Figure 1a NOTE Any loop name ending in Flo represents a certain gas e g AirFlo O2Flo etc with a dedicated TMFC If the gas loop name does not end with Flo it represents the presence of a gas solenoid valve rather than a TMFC The Gs3Flo 3 gauge screen...

Page 104: ... controller including but not limited to setpoints control modes alarms P I values and cascades are saved to the controller s memory You can save this data with a unique name using the Save As button see Step 2b in Figure 56 on the following page or overwrite an existing recipe using the Save button Step 2a in the same illustration Each controller is capable of storing up to 10 recipes You can ret...

Page 105: ... 1 Listed here are the current User Interface and Control Program versions To update the software see Section 16 3 2 To recalibrate the system s touchscreen press the Calib button then touch the onscreen target each time it appears You will be guided through the process 1 Press the recipe file of choice from the list in this box 3 Press the Load button to load the Selected Recipe file 2b If you wi...

Page 106: ...abinet 1 In the System Settings screen press the Refresh button to update the current software status and to search for a new USB drive 2 The name of the new drive folder appears in the Update File box 3 Press the Update button to install the file The file will reboot twice this may take a little time 4 The Software pane will reflect the changes Updating software will not affect any previous user ...

Page 107: ...is screen All new hardware scanned will appear in the New Hardware box see Figure 59 below 2 Press the button and the new hardware list will move into the Module box 3 Your BioFlo 415 needs a unique ID number in the SCADA pane assign the correct Communication Mode and Unit ID number then press the Set button Figure 59 Adding New Hardware Step 1 press here Step 2 press here ...

Page 108: ...e or button to move that user from one pane to the other An Administrator can also add users to or remove users from the system using the Add User and Remove User buttons To remove a user press the user name to select it in the pane where it appears then press the Remove User button To add a user press the Add User button use the keypad screen that opens see illustration on the following page to t...

Page 109: ...optic Summary and Trend navigation buttons remain active the other navigation buttons will be greyed out and inaccessible When the User Name and Password are entered as desired press the OK button When a user presses the User button that user can use the popup buttons as shown here to Log Off or to Log On by pressing the appropriate button If the user has Administrator status the Change Password b...

Page 110: ... 3 The pH and DO control loops have been set to AUTO mode The desired cascades have been enabled see Section 11 The pumps have been set to run and the liquid addition system is in place see Sections 16 1 16 2 Inoculate the vessel see Section 16 3 If you are using NBS supervisory software start BioCommand see Section 16 4 During the run To conduct sampling see Section 16 5 To review the various pha...

Page 111: ...p 2 Select the desired tubing size see Table 9 in Section 13 3 for reference and cut a length sufficient to reach from the inlet source through the pump and to the outlet recipient allowing a few extra inches 3 Form a loop large enough to go around the pump head 4 Pull the right side of the spring loaded lever down to release the tubing guide 5 Hold the upper spring loaded clip open and load the u...

Page 112: ...ils on pump set up and operation 16 2 Install Liquid Addition Systems Figure 61 on the following page is a simplified depiction of a typical addition system Depending on the liquids base acid nutrients media to be added your system may be slightly different 1 Aseptically install if applicable a sterile 0 2µm filter in one of the two penetrations on the addition bottle cap 2 Aseptically connect the...

Page 113: ...hey pass through the peristaltic pumps are critical parameters in the proper operation of a P I pH control system If the tubing is too large excessive doses will be added The result is that the system will overcontrol alternating in close succession between adding one liquid then the other providing little or no change in pH reading A user selected deadband value is an aid to control pH within the...

Page 114: ...lt Table 9 the flow rate tubing size chart for further information 16 3 Inoculation When you are ready to inoculate the media to start a batch observe aseptic techniques to inject the inoculum using a syringe through the self sealing rubber seal of the 12mm or 19mm port septum 16 4 Launch BioCommand if present Start BioCommand on your PC reset the EFT Elapsed Fermentation Time to zero and make app...

Page 115: ... sure the gasket remains in the cap 8 Holding the stem at its top remove the harvest sample port flange cap making sure the gasket remains in the groove on the port fitting 9 Aseptically insert the sampler needle into the the harvest sample septum until the sampler and port flanges are flush against each other 10 Orient the sampler as desired close the tri clamp over the connection to secure it 11...

Page 116: ...N EXHAUST OUT connection on the side of the cabinet 3 Following the sampling instructions provided in Section 16 5 above Sampling Procedure to harvest the entire contents of the vessel Note that a small residue will probably remain in the vessel Alternatively you can use a feed pump to harvest the vessel contents When the vessel is set up on the control cabinet adjust the level probe s tip to the ...

Page 117: ...ely this phase can last for a standard e coli fermentation for example approximately 2 3 hours Eventually however you must expect your batch to decline 16 7 4 Decline Phase This final phase is marked by a slow dying off It is of course inevitable 16 8 Batch Operation A batch operation is a closed growth environment in the sense that it contains a finite amount of media The inoculum grows through t...

Page 118: ...e likened to an assembly line process fresh medium is added as batch broth is harvested The fermentation vessel contains at all times the optimum amount of media with an established thriving culture Figure 64 is a simplified depiction of this setup Figure 64 Chemostat Operation Setup FRESH MEDIA FRESH MEDIA HARVESTED BROTH ...

Page 119: ...em follow these steps 1 Set GasFlo to OFF 2 Set Agit and Temp to OFF 3 Set all other control loops to OFF 4 Turn off the power 5 If the system is not to be used for several days disconnect the power plug 6 Remove drain and clean the vessel as outlined in Section 17 NOTE Never wash the filters or get them wet ...

Page 120: ...d microorganisms into the environment 2 Fill the vessel with tap water again up to or near the surface of the headplate 3 Keep Agitation at 200 300 rpm without aeration for about one hour at a temperature setpoint of 30 35 C 4 Drain the water while the Air Inlet valve is open set Gas flow rate at 5 10 L min then close it 5 Fill the vessel with tap water again up to or near the headplate and leave ...

Page 121: ...eadplate penetration O rings EPDM Metal surfaces 316L or 316 stainless steel Inoculation septum Pure gum rubber color tan 17 2 Cleaning the Cabinet At least once a month clean all the metal parts of your unit Use a soft damp cloth moistened with water or mild detergent If a detergent is used remove all residue by rinsing them with clean water ...

Page 122: ...th the electrode tip immersed in a solution of 3 molar KCl or a buffer solution between pH 4 00 and pH 7 00 CAUTION Never let a pH probe rest on its tip Never leave a pH probe in DI water 18 2 DO Probe Maintenance and Storage Use soft facial tissue to clean the DO probe Check the probe s Teflon membrane to be sure there are no punctures puckers or wrinkles If there are the probe should be replaced...

Page 123: ...eck that all controls and connectors are free of dust 3 Check that all O rings in the headplate are intact and in good condition Replace those that are not 18 5 Fuse Replacement There is one replaceable 5 Amp fast acting glass tube fuse 18 6 Port Adapter O Rings Each time you clean the vessel assembly check all port and adapter O rings for wear distortion rips or any other damage to their integrit...

Page 124: ...1360 0050 User s Guide 18 8 Replacement Parts See Section 19 2 Service for a list of replacement parts Please remember that using the NBS part number will facilitate processing of your order by your local NBS distributor ...

Page 125: ...You may be able to resolve the situation easily and quickly yourself If the problem is not listed below or if the suggested solutions do not work please call your NBS representative to request a service technician Other than the solutions proposed below do not attempt to fix the equipment yourself Problem Possible Solution TEMPERATURE Readout is a negative value typically 225 C Inspect the tempera...

Page 126: ...ration Probe may be defective replace it pH DO amplifier on superboard may be defective call for service GASFLOW There is insufficient gas flow Inlet or exhaust sterile air filter may be wet or clogged replace filter Check that the air pressure is within the specified range Make sure the control mode for DO and for pH is set to AUTO not OFF Make sure that the GasFlow loop is ON Make sure that the ...

Page 127: ...p Gasket 2 inch sight glass filter 2 P0280 1443 Tri Clamp Gasket inch EPDM 5 P0280 1444 Tri Clamp Gasket inch EPDM 5 P0280 2690 Septum single 5 P0280 5312 Sparge O Ring EPDM 5 P0280 5612 Drive O Ring EPDM 1 P0280 5652 Drive O Ring EPDM 1 P0280 5882 Baffle Plug O Ring EPDM 6 Harvest Port Adapter O Ring EPDM Sample Port Adapter O Ring EPDM Sparger Port Adapter O Ring EPDM Vessel Light O Ring EPDM P0...

Page 128: ...116 BioFlo 415 y M1360 0050 User s Guide 2 20 0 D DR RA AW WI IN NG GS S Figure 65 Controller Schematics overview ...

Page 129: ...117 New Brunswick Scientific User s Guide Figure 65a Controller Schematics left side ...

Page 130: ...118 BioFlo 415 y M1360 0050 User s Guide Figure 65a Controller Schematics right side ...

Page 131: ...5 Deleting a Control Loop 33 16 Deleting a Pump Control Loop 33 17 Calibration Screen 34 43 18 Cascade Screen 35 69 18a Cascade Screen detail 67 19 Trend Screen 35 72 20 Pump Screen also appears as Figure 43a 36 61 21 Alarm Summary Screen 36 82 22 Sterilization Screen 37 54 23 Controller Setup Screen 38 87 88 24 Recipe Manager Screen also appears as Figure 56 38 25 System Settings Screen 39 26 Har...

Page 132: ...ry Screen 84 50 Sample Alarms History Screen 86 51 Controller Setup Screen 0 1 TMFC 89 52 Controller Setup Screen 2 TMFCs 90 53 Controller Setup Screen 3 TMFCs 90 54 Controller Setup Screen 4 TMFCs 91 55 Gs3Flo 3 Gauge Screen 92 56 Recipe Manager Screen 93 57 System Settings Screen 93 58 Hardware Setup Screen 95 59 Adding New Hardware 95 59a Security Settings Screen 96 59b Security Keypad 97 59c U...

Page 133: ...es 29 3 Sterilization Sequence 37 4 AFS Modbus Com Port Pin Designation 41 5 Changing Sterilization Parameters 56 6 Valve Sequencing Labels Operations 57 7 Trend Graph Buttons 74 8 Pump Control Modes 78 9 Flow Rate per Tubing Size 79 10 Alarm Summary Screen Features 83 11 Wetted Parts 109 12 Contents of Spare Parts Kit M1360 9969 115 ...

Page 134: ...tuators that maintain the various parameters at their setpoints Using temperature as an example the controller compares the output of a temperature sensor to the user entered temperature setpoint and generates a signal to activate either a heater or a cooler to maintain vessel temperature at the temperature setpoint The controller provides the logic that generates appropriate drive signals to vari...

Page 135: ...oportional P integral I and derivative D terms The P I and D constants are three numbers that determine the relative sizes of the proportional integral and derivative terms respectively To use a temporal analogy the P or proportional part of the control signal reflects present deviations between setpoint and current value The I or integral component reflects past deviations and the D or derivative...

Page 136: ...his task A number of textbooks1 that explain the theory and describe the process could be useful for the mathematically inclined novice The Ziegler Nichols method described in the footnoted reference is used at our production facilities The following suggestions are intended for novices Be sure to refer to a textbook and consider utilizing the services of a technician Allow sufficient time for the...

Page 137: ...a normalizing constant for the loop The controller reevaluates SN every second I is divided by 60 so any value entered by the user should be in reciprocal minutes The normalizing constant k can be set to any non zero value but is usually set to the full scale reading of the loop For example if the range of expected temperatures is 0 to 125 setting k to 125 results in a P term value of P when the e...

Page 138: ...itation 1000 RPM Aeration 1 VVM 22 1 1 OTR Calculations OTR can be estimated by titrating a fixed amount of sodium sulfite Na2SO3 with air CU 2 2SO3 02 2SO4 The Procedure Calibrate the DO electrode Set zero on DO Fully oxygenate the fermentor with agitation and airflow Set span to 100 Introduce a known amount of Na2SO3 into the fermentor when fully oxygenated OTR 30 000 n mM O2 L hr V Δ T n number...

Page 139: ...power utilized by a given number of impellers the following formula can be used The impeller diameter varies to the 5th power with respect to horsepower A very slight change in the diameter of an impeller can make a great deal of difference in the HP required to drive that impeller The approximate horsepower utilized to drive a given set of impellers is determined as follows HP D5 x RPM3 x 4 5 x 1...

Page 140: ...ioFlo 415 the information is generally applicable for any fermentation 23 1 Media Formulation Question What kind of media should be used and does it differ from media used in shake flasks Answer The media used in shake flasks does differ from the standard media used in a fermentation vessel Shake flask media is generally of a much simpler composition LB Broth and Tryptic Soy Broth are standard sha...

Page 141: ... L When the foam probe is used the pumping of antifoam is controlled by the unit The pump should be set to add the minimum amount of antifoaming agent required to prevent foaming in your particular process That amount varies depending on the amount of protein in the media the amount of protein secreted by the microorganism agitation speed and other factors Therefore you will have to experiment to ...

Page 142: ...The needle will corrode but it saves the fermentor vessel Insert the needle though a septum port so that the drip point is away from stainless steel components and fairly close to the liquid level You may also use a more diluted solution of the acid or base However take note that this may cause the complication of adding a larger volume of liquid to the vessel Also it is not a good idea to add aci...

Page 143: ...ence 23 6 Recommended Process Control Settings Question What are the recommended process control settings i e temperature pH agitation speed DO gas sparge rate Answer For E coli temperature is usually set to 32 35 C and pH is set at 7 0 7 2 For yeast the values are 30 C and a pH value of 5 0 Agitation speed is usually set to a minimum of 200 300 rpm with a maximum value of 1000 rpm Dissolved oxyge...

Page 144: ...d from Corrosion Data Survey 1967 C F Percent Concentration in Water and 1974 Editions published by the National Association of Corrosion Engineers The influence of contaminants is probably the most important from a commercial standpoint Corrosive solutions are seldom found that will be free to all contaminates However the majority of these contaminants have no influence on corrosion but the ones ...

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Page 159: ...Good Resilience Rebound Good Abrasion Resistance Good Tear Resistance Fair Solvent resistance Poor Oil resistance Poor Low Temperature Usage 20 to 60 F 29 to 51 C High Temperature Usage to 350 F 177 C Aging Weather Sunlight Excellent Adhesion to Metals Fair to Good Ethylene Propylene is a polymer with outstanding properties It has exceptionally good weather aging and ozone resistance excellent wat...

Page 160: ...am Probe 6 B Backflow Prevention Kit 49 50 Base Concentration 130 Base Type 130 Batch Operation 105 BioCommand 7 22 40 C Cabinet Cleaning of 109 Calibration of Touchscreen 27 93 Calibration Screen 34 Cascade Creating a 67 Cascade Creating a 68 Cascade Screen 35 68 CAUTION Symbol for iv Cautions Recap of 1 Certifications 25 Chemostat Operation 106 Chemostat Operation Setup 106 CO2 4 89 CO2Flo 3 90 ...

Page 161: ...l 6 Front View 13 Fuse Replacement 111 G Gas Connections 12 Gas Quality Issues 54 GasFlo 89 Gauge Screen 34 Glucose Feed Concentration 130 Gs3Flo 3 91 H Harvesting 104 Headplate O Ring Replacement 111 Horsepower Factors that Affect 127 HOT SURFACE Symbol for iv I Index of Drawings 119 Index of Tables 121 Information Sheet vi Inoculation 102 Inputs Outputs 23 Inspection of Boxes 3 Installation Gas ...

Page 162: ...ations for 131 Pump Array Standard 77 Pump Assignment 61 Pump Assignment Screen 64 Pump Calibration 65 79 Pump Control Modes 78 Pump Flow Rate 79 Pump Period sec 80 Pump Setpoints 77 Pumps Screen 36 PWM 54 R Read Line 75 Recipe Loading a 92 Retrieving a 92 Saving a 92 Recipe Files 92 Regulatory Compliance 25 Remove User 96 Renaming Control Loops 31 Replacement Parts 112 Replacing Headplate O Ring ...

Page 163: ...xi Temperature Control 5 RTD 5 Setpoint 5 Touchscreen Calibrating the 27 93 Installing the 8 Trend Graph 73 Creating a 72 Trend Graph Buttons 74 Trend Screen 35 Trend Setup Screen 73 Troubleshooting 113 Tubing Recommendations 129 Tubing Size 129 U User Button 96 97 Utilities 10 V Valve Sequencing 55 56 Valve Sequencing Labels Operations 57 Vessel Description of 4 Vessel Approval 25 Vessel Pressuri...

Page 164: ...152 BioFlo 415 y M1360 0050 User s Guide ...

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