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NEW BRUNSWICK SCIENTIFIC BioFlo 415 Manual To Operations

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BioFlo 415
Benchtop SIP
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

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Summary of Contents for NEW BRUNSWICK SCIENTIFIC BioFlo 415

  • Page 1 BioFlo 415 Benchtop SIP 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 Fax: 49 (0)7022 32486 E-mail: sales@nbsnv-sa.be E-mail: sales@nbsgmbh.de CHINA THE NETHERLANDS New Brunswick Scientific Co., Inc. New Brunswick Scientific BV A903, 904 Yin Hai Building Kerkenbos 1101, 6546 BC Nijmegen No. 250, Cao Xi Road P.O Box 6826, 6503 GH Nijmegen Shanghai 200235, P.R.
  • Page 3 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.
  • Page 4 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.
  • Page 5 W A R R A N T Y 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...
  • Page 6 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: __________________________________________________ __________________________________________________ __________________________________________________ __________________________________________________ __________________________________________________ BioFlo 415 • M1360-0050 User’s Guide...

  • Page 7: Table Of Contents

    4.6.4 Install Motor Assembly..................18 4.6.5 Connecting Vessel to Cabinet ................18 ..................21 LECTRICAL ONNECTIONS ....................21 OWER WITCH ................22 PTIONAL OMMAND 4.10 ............. 22 NPUTS UTPUTS FOR NCILLARY EVICES SPECIFICATIONS ............................. 24 New Brunswick Scientific User’s Guide...
  • Page 8 8.2.3 Starting a Sterilization Sequence ..............53 8.2.4 Resetting Sterilization Parameters..............55 ....................56 ALVE EQUENCING ................58 TERILIZATION HASE NDICATION ..................58 BORTING TERILIZATION PREPARING FOR FERMENTATION ...................... 59 ....................... 59 YSTEMS HECK BioFlo 415 • M1360-0050 User’s Guide...
  • Page 9 Gas Control with 1 or No TMFC ..............89 15.1.2 Gas Control with 2 TMFCs ................89 15.1.3 Gas Control with 3 TMFCs ................90 15.1.4 Gas Control with 4 TMFCs ................91 15.2 ..................... 92 ECIPE ANAGER 15.3 ..................... 93 YSTEM ETTINGS New Brunswick Scientific User’s Guide...
  • Page 10 ....................113 ROUBLESHOOTING 19.2 ....................... 115 ERVICE ARTS DRAWINGS.............................. 116 20.1 ....................119 IST OF RAWINGS 20.2 ....................... 121 IST OF ABLES APPENDIX A: SOME GENERAL CONCEPTS..................122 21.1 ?..................122 HAT IS A ONTROLLER BioFlo 415 • M1360-0050 User’s Guide...
  • Page 11 ......................130 LUCOSE 23.6 ............131 ECOMMENDED ROCESS ONTROL ETTINGS APPENDIX D: CORROSION RESISTANCE TABLES................. 132 APPENDIX E: GENERAL CHARACTERISTICS OF EPR..............147 25.1 EPR ....................147 DENTIFYING 25.2 .................. 147 ENERAL HARACTERISTICS INDEX............................... 148 New Brunswick Scientific User’s Guide...

  • Page 13: Warnings & Cautions

    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 415. Please take a moment to acquaint yourself with the content of each message.
  • Page 14 Be sure to wear protective gloves when installing a glass electrode. We recommend that you avoid the use of hydrochloric acid (HCl) with the BioFlo 415 for pH control or any other purpose, because HCl corrodes stainless steel. Over time, it will severely damage the vessel, a costly component to replace, and other stainless steel components.

  • Page 15: Inspection & Unpacking Of Equipment

    & 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.

  • Page 16: Introduction & Overview

    & & System BioFlo 415 is a versatile sterilizable-in-place (SIP) fermentor that provides a fully equipped system in one compact package. It can be employed for batch or continuous culture with process control of pH, dissolved oxygen (DO), agitation, temperature, pump feed, antifoam, level and addition analog/digital inputs and outputs.

  • Page 17: Temperature Control

    Default P & I (proportional & integral) values are preset at the factory. We strongly recommend that you maintain the factory-set parameters. New Brunswick Scientific User’s Guide...

  • Page 18: Do Control

    The sampler assembly consists of the sampler tube with a sampler needle, three tri- clamps and a sampler valve. This assembly facilitates the aseptic removal of the sample, or harvesting the entire contents of the vessel, to an optional sample/harvest vial or bottle. BioFlo 415 M1360-0050 User’s Guide...

  • Page 19: Recommended Accessories & Supplies

    3.11 Recommended Accessories & Supplies Before you begin to assemble your BioFlo 415, it would be prudent to verify that you have all of the following accessories and supplies readily at hand: • Rubber gloves • Media • Silicone tubing •...

  • Page 20: Installation

    Ensure that the surface can bear the weight of the fermentor plus vessel contents and any applicable ancilliary equipment. Also ensure that there is enough space around the back and the front of the BioFlo 415 for proper operation and access. Allow at least 4 inches of clearance behind the unit for heat dissipation.
  • Page 21 Before making electrical connections, verify that the supply voltage matches 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. New Brunswick Scientific User’s Guide...

  • Page 22: Utilities

    (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. BioFlo 415 M1360-0050 User’s Guide...

  • Page 23: Electrical Requirements

    (located on the rear panel of the cabinet) and the control schematics supplied with the unit. WARNING! High voltage. Always make sure this equipment is properly grounded. New Brunswick Scientific User’s Guide...

  • Page 24: Water And Drain Connections

    This is needed because the gases leaving a fermentor are moisture-laden and filter condensate also exits through this connection. BioFlo 415 M1360-0050 User’s Guide...

  • Page 25: Vessel Assembly

    As you set up the BioFlo 415, the control cabinet must be positioned to the right of the vessel as you face the system, to facilitate connections. Position the cabinet first.
  • Page 26 DO/pH LEVEL 12mm SPARE PORT (SINGLE SEPTUM) FRONT of VESSEL 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. BioFlo 415 M1360-0050 User’s Guide...

  • Page 27: Sparge & Exhaust Assemblies

    This assembly (see Figure 4) consists of the sparge valve, sparge line, inlet filter and condensate return line. The assembly should be installed in one of the 12mm ports on the vessel headplate; see Figures 3a & 3b for the recommended location. New Brunswick Scientific User’s Guide...
  • Page 28 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). BioFlo 415 M1360-0050 User’s Guide...

  • Page 29: Other Ports & Plugs

    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 the probe is firmly attached. New Brunswick Scientific User’s Guide...

  • Page 30: Install Motor Assembly

    (see Figure 1a) to the air inlet filter housing on top of the sparger, on the vessel, using the tubing and tri-clamp connector supplied. Make sure that the tri-clamp gasket is in place before the tri-clamp is secured. BioFlo 415 M1360-0050 User’s Guide...
  • Page 31 FRONT of VESSEL WATER OUTLET To lower exhaust condenser water inlet WATER INLET 6. With reference to Figures 7 (above) and 5, connect the ’s water JACKET inlet “T” to the exhaust condenser’s lower connection. New Brunswick Scientific User’s Guide...
  • Page 32 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 that go to the drain line. Figure 1a: Control Cabinet Service Connections BioFlo 415 M1360-0050 User’s Guide...

  • Page 33: Electrical Connections

    Do not plug the heater cable into the cabinet unless the vessel jacket has been primed (see Section 8.2.1). Main Power Switch The main power switch is located on the righthand side of the control cabinet, below the touchscreen and above Pumps 1-3. New Brunswick Scientific User’s Guide...

  • Page 34: Optional Biocommand Plus

    A 25-pin RS232/422 Modbus com port is provided on the rear panel of the control cabinet (see Figure 8 on the following page) to connect the BioFlo 415 to a supervisory host computer (see also Section 6.3). Communications to BioCommand Plus software are via an optional RS-232 interface cable: 1.
  • Page 35 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. New Brunswick Scientific User’s Guide...

  • Page 36: Specifications

    BioFlo 415 System 19.5L Vessels Total Volume 2 - 5.25L 4 – 10.5L 5 – 15.5L Working Volume Aspect Ratio Controls 1 vessel with 32 control loops; stores 10 Controller Control Station recipes & 8 process variables per vessel for trend graphing.

  • Page 37: Certifications

    See Table 9 (Section 13.3) for pump flow rates according to tubing size Vessel weight does not include probes, exhaust condenser or other options. Certifications The BioFlo 415 has been tested to ETL standards, to comply with the safety standards: UL 61010A-1 UL 61010A-2-010 CAN/CSA-C22.2 No.
  • Page 38 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. BioFlo 415 M1360-0050 User’s Guide...

  • Page 39: Operating Controls

    Touchscreen Your primary interface with the BioFlo 415 is the touchscreen on the control cabinet. Figure 9: Touchscreen TOUCHSCREEN DISPLAY CONTROL CABINET ON/OFF POWER SWITCH PUMPS Display Screens 6.2.1 Touchscreen Calibration The first time you power up, you will be prompted to calibrate the screen to your touch.

  • Page 40: Start-Up Screen

    Start-Up Screen The Start-Up screen, which tells you which operating software version is installed in your BioFlo 415, is first screen you see each time you turn on the power, if you have already calibrated the touchscreen (see Section 6.2.1):...
  • Page 41 PHASE INDICATOR and ICON VESSEL LIGHT button: press once to turn light on for 10 minutes; press Your again to turn off BioFlo 415 comes with pre-assigned loop names. SCROLL UP As you add buttons more, use the SCROLL DOWN...

  • Page 42: Synoptic Screen

    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.

  • Page 43: Gauge Screens

    When adjusted, the scaling for the gauge will also be Decimal Places: adjusted to reflect Press the appropriate the high & low button to display limits selected. values with 0, 1, 2 or 3 decimal places. New Brunswick Scientific User’s Guide...

  • Page 44: Adding Loops

    6.2.6 Adding Loops The BioFlo 415 comes to you with standard factory-assigned loops and the possibility to add more loops, which are related to external auxiliary equipment, added via standard analog and optional serial (RS232) input/outputs located on the rear panel.

  • Page 45: Deleting Loops

    (such as Pump 4 screen where shown here) have a you can delete Settings button. the pump loop by pressing on the Remove button. SUMMARY 5. When you return to the screen, the loop will be deleted. New Brunswick Scientific User’s Guide...

  • Page 46: Selecting Loop Control Modes

    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. BioFlo 415 M1360-0050 User’s Guide...

  • Page 47: Cascade Screen

    The data can be exported through the USB port in Excel format to a PC. For details on using the TREND screen, see Section 12. Figure 19: Trend Screen The user will assign a tracking color to each loop. New Brunswick Scientific User’s Guide...

  • Page 48: Pump 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 Screen BioFlo 415 M1360-0050 User’s Guide...

  • Page 49: Sterilization Screen

    The sterilization sequence used is based upon the standard sterilization methods currently used by New Brunswick Scientific. This sequence uses the following parameters to automatically control the heating and cooling of the vessel: Table 3: Sterilization Sequence...

  • Page 50: Setup Screen

    This master screen (see Figures 23, 24, 25 & 26) is actually comprised of four screens, accessed by tabs, which are used to set up the controller, recipe management, system settings and hardware for the BioFlo 415 system. This section will introduce you to those screens and their features; for details on the SETUP screen, including a fifth tab that may be present, see Section 15.

  • Page 51: Rs232/422 Computer Interface

    An RS232/422 com port has been provided; there is a 25-pin “D” connector located on the lower rear panel of the control cabinet (see Figure 8, repeated on the following page for easy reference). It is labeled AFS/Modbus New Brunswick Scientific User’s Guide...
  • Page 52 Table 4, on the following page, identifies the pin designations for this 25-pin RS232/422 connector: BioFlo 415 M1360-0050 User’s Guide...
  • Page 53 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 address for the machine is 8. New Brunswick Scientific User’s Guide...

  • Page 54: Probe Preparation & Calibration

    4. Rinse the pH electrode with distilled water, then immerse it into pH 7.00 buffer solution and allow a few minutes for the system to equilibrate. 5. Open the CALIBRATION screen (see Figure 17, repeated below for easy reference). Steps 6-8 are indicated as callouts around this screen: BioFlo 415 M1360-0050 User’s Guide...
  • Page 55 Take a sample from the vessel and compare the pH value displayed on the control cabinet screen to the pH recorded by an external SET ZERO pH meter. Any discrepancy should be adjusted with the procedure. New Brunswick Scientific User’s Guide...

  • Page 56: Ph Probe Installation

    Also make sure that there is no interference inside the vessel. Figure 27: pH Probe Installation (gel-filled) pH PROBE O-RINGS HEADPLATE BioFlo 415 M1360-0050 User’s Guide...

  • Page 57: Ph Probe Maintenance & Storage

    CAUTION! We recommend that you avoid the use of hydrochloric acid (HCl) with the BioFlo 415 for pH control or any other purpose, because HCl corrodes stainless steel. Over time, it will severely damage the vessel, a costly component to replace, and other stainless steel components.

  • Page 58: Do Probe Preparation

    If you are using an adapter for a 19mm port: 1. Screw the adapter into the port until it is finger-tight. 2. Carefully insert the probe into the adapter. 3. Finger tighten enough to compress the O-ring to ensure a tight seal. BioFlo 415 M1360-0050 User’s Guide...

  • Page 59: Do Probe Polarization

    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: Method 1: 1. Remove the DO cable from the DO electrode. New Brunswick Scientific User’s Guide...

  • Page 60: Do Probe Calibration: Setting Span

    10 minutes (this may take up to 30 minutes total). 4. In the CALIBRATION screen, select (see Figure 29). 5. Enter 100 in the SET SPAN edit box, then press the SET SPAN button. BioFlo 415 M1360-0050 User’s Guide...

  • Page 61: Automatic Sterilization

    Automatic Sterilization is activated at the touchscreen interface. The controller is preconfigured for this feature. WARNING! To avoid possible safety hazard, the BioFlo 415 Backflow Prevention Kit, NBS PN M1360-5002, must be installed in the water inlet line before performing a sterilization cycle! Be sure to follow the instructions in Section 8.1 to properly install the kit...

  • Page 62: Bioflo 415 • M1360

    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 1-732-287-1200 or Bioinfo@nbsc.com. BioFlo 415 M1360-0050...

  • Page 63: Sterilization

    Make allowances for inoculum to be added later. Replace the fill port cap and tighten it securely. NOTE Allow for a loss of up to one liter of media during the sterilization cycle. New Brunswick Scientific User’s Guide...

  • Page 64: Priming The Water Jacket

    2. Set the temperature loop control mode to 3. Check that the temperature reading is higher than 5°C. 4. Allow water to enter the piping system; it will stop at solenoid valve TY- 5. Set the temperature control mode to AUTO BioFlo 415 M1360-0050 User’s Guide...

  • Page 65: Sterilization Safety Features

    150 - 300 RPM. Turn on the Agitation control loop by putting the loop in control. AUTO 5. Press the button to open the STERILIZATION screen (see Figure Sterilize 22, repeated on the following page for easy reference). New Brunswick Scientific User’s Guide...
  • Page 66 WARNING! All metal surfaces remain very hot after sterilization. Be sure to wear protective gloves when you move the sparge valve from sterilization to sparge position. BioFlo 415 M1360-0050 User’s Guide...

  • Page 67: Resetting Sterilization Parameters

    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 follows the illustration. Figure 31: Automatic Sterilization Parameters New Brunswick Scientific User’s Guide...

  • Page 68: Valve Sequencing

    Valve Sequencing The controller has been preconfigured to sequence a set of valves on the BioFlo 415 during the automatic sterilization cycle. The column headings or operations for valve sequencing are as explained in Table 6 on the following page: BioFlo 415 M1360-0050 User’s Guide...
  • Page 69 TY-5K, white. and Exhaust Heater Growth settings are black & white. The legend for these settings is shown in Figure 33 on the following page, which is also a detail from the STERILIZATION screen: New Brunswick Scientific User’s Guide...

  • Page 70: Sterilization Phase Indication

    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. BioFlo 415 M1360-0050 User’s Guide...

  • Page 71: Preparing For Fermentation

    Systems Check Prior to any fermentation run, make sure that all standard and optional systems provided with the BioFlo 415 are operational: • Exhaust Heater • Agitation Control • pH Control • Temperature Control • Level 1 Control • DO Control •...

  • Page 72: Adding Media To The Vessel

    Please allow for a loss of up to one liter of media during sterilization. 5. Once the media (or PBS) has been added, the port used must be closed, along with any other open port on the headplate. BioFlo 415 M1360-0050 User’s Guide...

  • Page 73: Getting Started

    Figure 20: Pump Screen NOTE: If there are more than 3 pumps, use these >>> and <<< buttons to 2. Press the navigate between pump’s screens. ASSIGNMENT button to open the Assignment pane (see Figure 35). New Brunswick Scientific User’s Guide...

  • Page 74: Settingp & I Values

    Setpoint box for the desired loop on the SUMMARY SYNOPTIC screen. In this example, we have selected Agit 2. The loop GAUGE screen opens (see Figure 13, repeated on the following page for easy reference): BioFlo 415 M1360-0050 User’s Guide...

  • Page 75: Modifying Setpoints

    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. New Brunswick Scientific User’s Guide...

  • Page 76: Using Level Probes To Program Feed Pumps

    When the vessel is set up at the control cabinet, raise the probe to the level at which you want harvesting to begin (i.e., above the current liquid level). Never lower a probe after sterilization! BioFlo 415 M1360-0050 User’s Guide...

  • Page 77: Level Control Off

    Section 10.6 for details). NOTE: To assure the most accurate flow rate, calibrate the pump (see Section 13.3) each time you change tubing. New Brunswick Scientific User’s Guide...

  • Page 78: Exhaust Heater

    10.6.5 Exhaust Heater The BioFlo 415 has been provided with a heater band on the exhaust line. The purpose of the heater is to prevent the exhaust filter from becoming saturated with condensate. To activate the heater, do the following: If 1 TMFC is installed: 1.

  • Page 79: Cascade Control

    PID-based control algorithm. The BioFlo 415’s RPC controller allows cascading from any loop to as many as five other loops. DO and pH are the most commonly cascaded-from loops; oxygen and nitrogen...
  • Page 80 When the PV equals the setpoint, the Out% should be approximately 0, as the controller senses no need to make adjustments to DO: To create a cascade: CASCADE CASCADE 1. Press the button to open the screen (see Figure 18, repeated here for easy reference): BioFlo 415 M1360-0050 User’s Guide...

  • Page 81: Controlling Do By Cascade

    @DO End Out% is reached. the cascade. Typically this value is the maximum allowable value the process can handle. In this example, when the DO End Out% reaches 25, the agitation will have increased to 1000 rpm. New Brunswick Scientific User’s Guide...
  • Page 82 DO PV are close to equal at the start of a run, it will work best if the cascade DO Start Out% is 0. When configured this way, any drop in DO below the setpoint will be compensated by the cascade loop. BioFlo 415 M1360-0050 User’s Guide...
  • Page 83 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. New Brunswick Scientific User’s Guide...

  • Page 84: Plotting Trends

    1. From any screen, press the button to open the screen (see Figure 19, repeated below for easy reference). Figure 19: Trend Screen 2. To select the first loop you wish to display, press the red Setup: button. BioFlo 415 M1360-0050 User’s Guide...
  • Page 85 Trend Graph buttons at the bottom of the graph: Figure 39: Trend Graph Touching the PV, SP or OU box will open the loop gauge screen (see Section 6.2.5). TREND GRAPH BUTTONS RAMP DOWN RAMP UP New Brunswick Scientific User’s Guide...

  • Page 86: Using The Export Button

    5. Press the Save button to save the file to the USB external memory device. 6. Remove the USB external memory device and use it to download the data to your PC. BioFlo 415 M1360-0050 User’s Guide...

  • Page 87: Using The Zoom Button

    (see Figure 41 on the following page). 3. The Read Line time is displayed at the bottom of the graph. New Brunswick Scientific User’s Guide...
  • Page 88 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. BioFlo 415 M1360-0050 User’s Guide...

  • Page 89: About Pumps

    1. Open the Pump Screen . Gauges for Pumps 1-3 are displayed in this screen (see Figure 43). If you have one or more additional pumps, press the >>> button to continue past Pump 3. New Brunswick Scientific User’s Guide...

  • Page 90: Pump Control Mode

    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. BioFlo 415 M1360-0050 User’s Guide...

  • Page 91: Pump Flow Rate & Calibration Methods

    Amount pump will start running. Pumped edit box. 9. Press the Set button to save this data to the PUMPS screen. NOTE: Calibration must be performed at operating setpoint. New Brunswick Scientific User’s Guide...

  • Page 92: Pump Period

    When selecting an external pump to operate with your system, please consult with your local sales representative to be sure the model you choose is compatible with your BioFlo 415. 1. Connect the D connector of the pump’s 3-wire cable at Interface on the rear panel of the pump (see Figure 46).
  • Page 93 2. Locate the Analog Output Connections on the rear of the BioFlo 415 cabinet (see Figure 47). Figure 47: Rear Panel of BioFlo 415 Cabinet Dip Switches: 1-3 for mA or V provided to switch connectors 1-3 between mA (down)

  • Page 94: About Alarms

    Figure 21: Alarm Summary Screen 2. Press the first loop for which you want to enable an alarm. That loop’s individual Alarms Screen will open. For this example, we use the Agitation loop. BioFlo 415 M1360-0050 User’s Guide...
  • Page 95 3a. If you wish to 3b..and enter an set an Absolute Absolute High Llimit alarm, enter an here. Absolute Low Llimit here... 4b..and here to 4a. Press here to enable the Audible enable the Visual alarm. alarm... New Brunswick Scientific User’s Guide...

  • Page 96: Acknowledging An Alarm

    Figure 49: Current Alarms Summary Screen 4. Press the Index box for the alarm you wish to acknowledge. It will turn green. 5. Press the Acknowledge button. The alarm will be deleted from the screen. BioFlo 415 M1360-0050 User’s Guide...

  • Page 97: Alarms History

    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 an alarm is triggered, whether Visible and/or Audible, the controller records the event.
  • Page 98 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. <<< BioFlo 415 M1360-0050 User’s Guide...

  • Page 99: Using The Setup Screen

    Hardware Setup see Section 15.3), and to check or change the (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...
  • Page 100 Name your unit using the Unit Name edit box touchpad; a corresponding colored button tab. In this sample screen (Figure 23), the Unit Name, BioFlo 415, is also written on the menu line button tab. You may prefer to name the unit by its...

  • Page 101: Gas Control With 1 Or No Tmfc

    (see Figure 52 on the following page). SUMMARY The gas process loops you will find in the screen are labeled AirFlo (1) and O2Flo (2). The loops’ numbers, 1-2, correspond to the gas connections on the cabinet (see Figure 1a). New Brunswick Scientific User’s Guide...

  • Page 102: Gas Control With 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). BioFlo 415 M1360-0050 User’s Guide...

  • Page 103: Gas Control With 4 Tmfcs

    TMFC. The Gs3Flo (3) gauge screen (see Figure 55 on the following page) allows you to set parameters for the TMFC that controls this gas. New Brunswick Scientific User’s Guide...

  • Page 104: R Ecipe M Anager

    Recipe Manager screen, where all saved recipes are listed in the Available Recipes pane. Select the desired recipe by following Step 1 in Figure 56, then load it as shown in Step 3. BioFlo 415 M1360-0050 User’s Guide...

  • Page 105: Recipe

    System Settings screen (see Figure 57). Use this feature select the onscreen language you prefer, to reset the date and/or time, to update the software, and to calibrate the BioFlo 415 touchscreen. Figure 57: System Settings Screen Other languages are not available at this time.

  • Page 106: Resetting Date/Time

    4. The Software pane will reflect the changes. Updating software will not affect any previous user settings. 15.4 Hardware Setup The BioFlo 415 system you purchase is preset in the factory as “Unit1” with all the accompanying hardware. Press the fourth tab in the SETUP...
  • Page 107 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 button.

  • Page 108: Security Settings

    15.5 Security Settings The security feature on the BioFlo 415 provides two user access levels: Operators have access to routine operations but they cannot change loop operating modes or change security settings. Administrators have access to all operations including defining new users (operators and administrators) and setting security parameters.
  • Page 109 ADMIN is the default password. NOTE: When Security is enabled, only the Synoptic/Summary and Trend navigation buttons remain active; the other navigation buttons will be greyed out and inaccessible. New Brunswick Scientific User’s Guide...

  • Page 110: Operation

    After becoming familiar with the components and operation of each of the systems, subsystems, and options that are included on your BioFlo 415, you are ready to set up the fermentor/bioreactor for a run. NOTE When preparing the fermentor/bioreactor for a run, always make sure that all hardware is set up to control the process prior to entering any setpoints or activating the system at the controller.
  • Page 111 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 upper clip channel; allow the upper clip to close over the tubing. New Brunswick Scientific User’s Guide...

  • Page 112: Install Liquid Addition Systems

    4. Connect the tubing, securing it with a plastic tie, to the appropriate addition port on the headplate. 5. Remove the clamp. CAUTION! Proper pH control is critically dependent on tubing size, which should be as small as possible. Consult Table 9 for guidance. BioFlo 415 M1360-0050 User’s Guide...

  • Page 113: Addition Tubing Size

    Addition Tubing Size pH can be controlled by automatic additions of liquid acid and base. Additions are triggered by the BioFlo 415 controller, which is constantly comparing current pH value with the pH setpoint and making adjustments as necessary. The concentrations of acid and base, and the inner diameter of the acid and base addition tubing (where they pass through the peristaltic pumps), are critical parameters in the proper operation of a P&I pH control system.

  • Page 114: Inoculation

    2. Wrap the sampler assembly in an autoclavable cloth, foil or paper. 3. Sterilize the assembly in the autoclave for 20 minutes at 121°C. BioFlo 415 M1360-0050 User’s Guide...
  • Page 115 2. Turn on gas flow into the vessel to create some backpressure. 3. Allow the desired amount of product to flow into the sample container. 4. Observe aseptic techniques to remove the container and to cap it. 5. Immediately replace the container with another sterile receptacle. New Brunswick Scientific User’s Guide...

  • Page 116: Harvesting

    Lvl2 Wet , it will harvest as much as possible. 16.7 Fermentation Phases In a typical fermentation run, you can expect to see four characteristic phases: Lag, Exponential Growth, Steady State, and Decline. BioFlo 415 M1360-0050 User’s Guide...

  • Page 117: Lag Phase

    A fed batch operation includes the addition of media to feed the batch fresh nutrient and to dilute any build-up of toxic by-products in the broth, thereby extending the life and growth of the desired product. Figure 63 on the following page is a simplified depiction of this setup: New Brunswick Scientific User’s Guide...

  • Page 118: Chemostat Operation

    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 HARVESTED FRESH BROTH MEDIA BioFlo 415 M1360-0050 User’s Guide...

  • Page 119: Shutdown Procedure

    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. New Brunswick Scientific User’s Guide...

  • Page 120: Cleaning

    List of Wetted Parts For further reference in your choice of cleaning detergents, Table 11 (on the following page) provides a list of wetted parts in the vessel assembly and the materials they are made of: BioFlo 415 M1360-0050 User’s Guide...

  • Page 121: 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. New Brunswick Scientific User’s Guide...

  • Page 122: Maintenance

    Preventive maintenance keeps your equipment in proper working condition. When performed routinely, maintenance results in longer life for your equipment. It also reduces time lost due to equipment failure. WARNING! Always turn your BioFlo 415 off and disconnect the power cord before performing maintenance. 18.1 pH Probe Maintenance and Storage The pH probe should be stored standing upright, with the electrode tip immersed in a solution of 3 molar KCl or a buffer solution between pH 4.00 and pH 7.00.

  • Page 123: Vessel & Tubing

    1. Pull the old O-ring out with a knife or a flat-bladed screwdriver. 2. Lubricate the new O-ring with a light coat of silicone grease. 3. Insert the new O-ring, making sure it is well-seated in the groove. New Brunswick Scientific User’s Guide...

  • Page 124: Replacement Parts

    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. BioFlo 415 M1360-0050 User’s Guide...

  • Page 125: Service

    If any problems occur with your BioFlo 415 system or its individual components, do not attempt to perform any service on it. Unauthorized servicing may void the warranty. Please contact your local NBS Service Department or your local NBS distributor.
  • Page 126 Lower the pressure and restart sterilization cycle. GENERAL: • Touchscreen is not responding. Calibrate touchscreen. • Pressure builds in vessel and Exhaust filter is blocked. Replace filter and verify that flow goes to Zero. exhaust heater is on. BioFlo 415 M1360-0050 User’s Guide...

  • Page 127: Service Parts

    Level Probe Port Adapter O-Ring EPDM P0280-6742 Multi-Septum Port Adapter O-Ring EPDM P0280-8512 O-Ring for Vessel Headplate P0280-8612 O-Ring for Vessel Headplate P0740-2720C3 ¼-inch Silicone Tubing P0740-2721C3 3/8-inch Silicone Tubing S2114-8061 Screw for Vessel Shroud, 8-32 x ⅜ New Brunswick Scientific User’s Guide...

  • Page 128: Drawings

    Figure 65: Controller Schematics (overview) BioFlo 415 M1360-0050 User’s Guide...
  • Page 129 Figure 65a: Controller Schematics (left side) New Brunswick Scientific User’s Guide...
  • Page 130 Figure 65a: Controller Schematics (right side) BioFlo 415 M1360-0050 User’s Guide...

  • Page 131: List Of Drawings

    Calibration Screen (DO) Water Pressure Regulator Kit NBS Backflow Prevention Kit Moving Sparge Valve for Sterilization Automatic Sterilization Parameters Sterilization Sequence Settings Legend for Sterilization Sequence Settings Sterilization Phase Diagram Pump Assignment Screen 62, 64 ...continued... New Brunswick Scientific User’s Guide...
  • Page 132 77, 99 Setting Pump Setpoint Calibrating the Pump Flow Rate Pump Period(sec) Variable Speed Pump Rear Panel of BioFlo 415 Cabinet Sample Loop Alarms Screen (Agit) Current Alarms Summary Screen Sample Alarms History Screen Controller Setup Screen (0-1 TMFC) Controller Setup Screen (2 TMFCs)

  • Page 133: List Of Tables

    Sterilization Sequence AFS/Modbus Com Port Pin Designation Changing Sterilization Parameters Valve Sequencing Labels/Operations Trend Graph Buttons Pump Control Modes Flow Rate per Tubing Size Alarm Summary Screen Features Wetted Parts Contents of Spare Parts Kit (M1360-9969) New Brunswick Scientific User’s Guide...

  • Page 134: Appendix A: Some General Concepts

    Based on information from a sensor, the controller generates an actuator control signal that maintains a parameter at its setpoint. Control will fail if any element in the control loop fails. BioFlo 415 M1360-0050 User’s Guide...

  • Page 135: What Is Probe Calibration

    If the constants are too small, control response will be slow, and setpoints may never be reached. Again, this can be mistaken for defective components. P-I-D constants are usually established by methodical trial and error. New Brunswick Scientific User’s Guide...

  • Page 136: What Is P-I-D Tuning

    • The magnitude of the control signal depends on the P, I and D constants. It also depends inversely on a Normalizing Constant. For example, Chinks, F.G., Process Control Systems: Application, Design, and Tuning, McGraw-Hill (1988), New York, Auckland, Bogota, London, Toronto, Sydney, Tokyo, Montreal. BioFlo 415 M1360-0050 User’s Guide...

  • Page 137: What Do The Constants Mean

    0 to 125, setting k to 125 results in a P term value of P when the error is at a maximum, i.e.: /k) = P(125/125) = P Similarly, with a full-scale error, the I term (after 1 minute) and the D term will be I and D respectively. New Brunswick Scientific User’s Guide...

  • Page 138: Appendix B: Otr

    • OTR = 30,000 n mM O /L/hr V Δ T = number of moles of sodium sulfite = vessel volume in liters Δ T = time taken from DO curve at two points of 50% DO min. BioFlo 415 M1360-0050 User’s Guide...

  • Page 139: Some Factors That Affect Otr And Horsepower

    350 mM O /L/hr of OTR. The BioFlo 415 fermentor is supplied with two or three (19L) properly sized Rushton Impellers. Placement of the impellers should be as indicated in Figure 2. In some processes, users may wish to use a third impeller. Should this be the case, however, a smaller impeller diameter is required, since the systems are specifically designed such that the vessel diameter, motor, impellers, to produce a specific OTR.

  • Page 140: Appendix C: Fermentation Techniques

    It also addresses the preparation, autoclaving and clean-up procedures for the vessel and accessories. While this example refers specifically to an E. coli fermentation in a BioFlo 415, the information is generally applicable for any fermentation. 23.1 Media Formulation...

  • Page 141: Antifoam Formulation

    BioFlo fermentors. However, Marprene tubing may be used as well, as long as the tubing size does not exceed 3/16” bore x 1/16” wall. Marprene tubing of this size or smaller can be used with Watson-Marlow 101 pump heads under low pressure and with clockwise rotation. New Brunswick Scientific User’s Guide...

  • Page 142: Acid & Base

    This operation can be ® controlled automatically by BioCommand Plus, NBS' proprietary Windows -based software. BioFlo 415 M1360-0050 User’s Guide...

  • Page 143: Recommended Process Control Settings

    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. New Brunswick Scientific User’s Guide...

  • Page 144: Appendix D: Corrosion Resistance Tables

    Stress relieved Vapor harmful < 60 psi Ignites Organics > 2.25% Mo Fuming liquid over 400ºC Hygroscopic 300 psi Liberates HCL in water No SO Narcotic Volatile Hazardous under pressure Ignites combustibles Exothermic with water BioFlo 415 M1360-0050 User’s Guide...
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  • Page 159: Appendix E: General Characteristics Of Epr

    350 (177°C). Ethylene Propylene is a polymer where oil and solvent resistance is poor, however, it is fairly good in ketones and alcohols. It is not recommended for exposure to aromatic hydrocarbons. New Brunswick Scientific User’s Guide...

  • Page 160: Index

    Symbol for, iv Backflow Prevention Kit, 49, 50 Base Concentration, 130 Base Type, 130 Deadband, 5, 34, 101 Batch Operation, 105 Decline Phase, 105 BioCommand, 7, 22, 40 Deleting Loops, 33 Description of Vessel, 4 BioFlo 415 M1360-0050 User’s Guide...
  • Page 161 Gas Connections, 12 Maintenance Inspections, 111 Gas Quality Issues, 54 Manual Conventions, iv GasFlo, 89 Mass Flow Controller, 5 Gauge Screen, 34 Media Formulation, 128 Glucose Feed Concentration, 130 Motor Assembly Gs3Flo (3), 91 Installation of, 18 New Brunswick Scientific User’s Guide...
  • Page 162 Shutdown, 107 Probes Sparge Assembly, 15 Installing the, 17 Specifications, 24 Process Control Settings Start-Up Screen, 28 Recommendations for, 131 Steady-State Phase, 105 Pump Array Sterilization, 50 Standard, 77 Aborting, 58 Pump Assignment, 61 Enabling, 58 BioFlo 415 M1360-0050 User’s Guide...
  • Page 163 Trend Setup Screen, 73 Troubleshooting, 113 Tubing Recommendations, 129 Tubing Size, 129 User Button, 96, 97 Utilities, 10 Valve Sequencing, 55, 56 Valve Sequencing Labels/Operations, 57 Vessel Description of, 4 Vessel Approval, 25 Vessel Pressurization, 12 New Brunswick Scientific User’s Guide...
  • Page 164 BioFlo 415 M1360-0050 User’s Guide...

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