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Applikon ez-Control Operator's Manual

For in situ sterilizable bioreactors
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ez-Control for
In Situ sterilizable Bioreactors
OPERATOR MANUAL
Project 310420; PS 70L, IST (AU)

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Summary of Contents for Applikon ez-Control

  • Page 1 In Situ sterilizable Bioreactors OPERATOR MANUAL Project 310420; PS 70L, IST (AU)

  • Page 2: Table Of Contents

    OPERATOR MANUAL CONTENTS ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) TABLE OF CONTENTS GENERAL SAFETY ..........................4 Safety Symbols ..........................4 Safety Warnings ..........................5 GENERAL ..............................7 Introduction ............................7 System Overview ..........................7 Home Screen .............................8 Icons ..............................10 Keypad and Keyboard ........................11 Bar Graph and Trend View Screens ....................12...
  • Page 3 Emergency Stop During Operation ....................43 © 2015 Applikon-Biotechnology, Delft. All rights reserved. This document is protected by copyright law. It may not be reproduced, stored or made public in any form by any means without the prior written approval of Applikon Biotechnology B.V.

  • Page 4: General Safety

    OPERATOR MANUAL SAFETY ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 1 GENERAL SAFETY SAFETY SYMBOLS The following symbols are used on the equipment and in this manual. WARNING Risk of personal injury or danger to life.

  • Page 5: Safety Warnings

    WARNING Risk of physical hand injury caused by pumps. • In most cases, one or more pumps are installed in the ez-Control. • According to the European Community legislation on machinery, a pump drive is a machine with potential hazard. Only apply these drives with the pump heads and tubing that are described in this manual (chapter: pumps)! •...
  • Page 6 Verify the tube quality inside the pump head before every fermentation run. Additional information • Do not apply an Earth Leakage Circuit Breaker in the power supply of the Applikon control equipment (in an industrial environment or laboratory, an earth leakage circuit breaker is not mandatory). •...

  • Page 7: General

    A wet part for temperature control and steam sterilization. • An ez-Control with Power Box for measurement and control of process variables (pH, temperature, DO, level, pressure, and stirrer speed) with corresponding controller outputs in order to keep process conditions on set-point.

  • Page 8: Home Screen

    Project 310420; PS70 IST (AU) 2.3 HOME SCREEN After switching on the power of the ez-Control, it presents itself (after initialization) on the touch screen display with its Home Screen.This will be either the Synoptic Screen or the Bar Graph Screen.
  • Page 9 (not a button) or a button without access. Colours of buttons Depending on the status of the ez-Control, the specific control loop and the actual screen and on the login level, the Parameter buttons and other functional (control) buttons have different colours.

  • Page 10: Icons

    High Alarm Limit Low Alarm Limit Upper Range Value Lower Range Value Applikon logo: enables changing the Project name / Application name Login (Access Control) Logout (returns to the View level) Upper Limit of the Actuator action < 100 % (default = 100 %) Low Limit of the Actuator action >...

  • Page 11: Keypad And Keyboard

    Project 310420; PS70 IST (AU) 2.5 KEYPAD AND KEYBOARD The ez-Control presents a numeric keypad when data like Set-points and parameter values must be edited. Follow the instructions shown above the edit field. The current value of the parameter is shown in a blue edit field. During editing, this field has a white background.

  • Page 12: Bar Graph And Trend View Screens

    Project 310420; PS70 IST (AU) 2.6 BAR GRAPH AND TREND VIEW SCREENS The Bar Graph screen is an overview of the various parameters of the ez-Control. Each Set-point value that is shown on a blue Set-point Value button can be edited (see Section 2.7).

  • Page 13: Set-Points

    OPERATOR MANUAL GENERAL ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) The Trend View screen shows the following information as a function of time: • Two red lines represent the alarm limits. • A black line shows the measured process value.

  • Page 14: Configurable Outputs

    4.3 (System Configuration) of this manual! 2.9 RELATED MANUALS • ez-Control Hardware Manual for In Situ sterilizable Bioreactors, • ez-Control Operator Manual for In Situ sterilizable Bioreactors, • ez-Control PID Control Manual for In Situ sterilizable Bioreactors, • ez-Control Extended I/O Manual.

  • Page 15: General Preparations

    Before mounting the sensor in the DN 25 sensor holder in the reactor, it must be calibrated. Preparation Connect the pH sensor and the temperature sensor to the right-hand side of the ez-Control. Start the ez-Control. On the touch screen: Press the pH parameter button.

  • Page 16: Check The Do Sensor

    After a stability check, the second calibration value is also stored and the calibration data (slope and offset) are calculated and stored in the memory of the ez-Control. For additional information concerning the pH calibration, see the ez-Control Software Reference manual, section “pH Calibration Routine”.

  • Page 17: Prepare Supplies

    OPERATOR MANUAL GENERAL PREPARATIONS ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Rupture disc If applied, verify that the rupture disc is free of cracks and properly fitted. Filling the reactor Fill the reactor with water or medium to working volume (only in case the reactor is sterilized while filled with liquid).

  • Page 18: Pressure Test And Sterilization

    Project 310420; PS70 IST (AU) 4 PRESSURE TEST AND STERILIZATION The pressure hold test and sterilization routine is fully controlled by the Software of the ez-Control (see the ez-Control Software Reference Manual, section “Menu Options”). The routine consists of a sequence of discrete steps.
  • Page 19 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Step 1: Start (full or empty) sterilization At this point in the procedure, the operator needs to confirm that the pH-sensor is calibrated. The states of the valves and other related actuators are kept as they were during fermentation / cultivation.
  • Page 20 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Step 4: Phase 1 Sterilization is activated: the temperature set-point is set to sterilization temperature (e.g. 123 °C). Steam valves are opened. Steam is introduced in the reactor jacket. Water (and condensate) from the jacket of the reactor is drained.
  • Page 21 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Step 5: Preheating Filter When the reactor temperature rises above a defined temperature, the Steam-to-filter valve (V-10.15) is opened. As a result, the Gas Inlet filter can be preheated with steam.
  • Page 22 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Step 6: Phase 2 After pre-heating the filter, steam is now injected directly into the reactor through the gas inlet filter: sparger valve V-10.08 must be fully open and overlay V-10.07 must be opened in such a way that a steam flow through the sparger is realized.
  • Page 23 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Step 8: Natural cooling (for Empty Sterilization only) Steam inlet valves are closed. In order to prevent vacuum in the reactor, the Gas inlet valve is opened but no cooling water is introduced into the jacket of the reactor yet.
  • Page 24 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Step 9: Cooling 2 In case of full sterilization, the Natural Cooling step is omitted; Gas Inlet Filter Condensate Valve V-10.10 is closed and Off-gas Filter Condensate Valve V-11.10 must be opened slightly in order to remove any condensate.
  • Page 25 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Circulation during cultivation or fermentation During cultivation, water is circulated through the jacket and the reactor is aerated through the sparger and overlay assembly.

  • Page 26: Sterilization Settings

    OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 4.3 STERILIZATION SETTINGS The sterilization settings for empty and full sterilization are presented in the tables below: EMPTY STERILIZATION FULL STERILIZATION Duration Temp.

  • Page 27: Actuator Activity Matrix

    OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 4.4 ACTUATOR ACTIVITY MATRIX Table Legend: In the tables below (empty and full sterilization) the following abbreviations and colors are used: = OPEN or ON...
  • Page 28 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) tuned to such extent that the gas/steam flow is equally divided between Sparging and Overlay! • During Phase 2 and Sterilization, Off-gas Block valve V-11.02 must be tuned to such extent that the reactor pressure can reach 1.2 ..
  • Page 29 OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) • From the Preheating Filter interval to the Natural Cooling phase, Condensate valve V-10.10 must be slightly open to drain the condensate from the filter housing.

  • Page 30: Execution Of The Sterilization Routine

    OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 4.5 EXECUTION OF THE STERILIZATION ROUTINE 4.5.1 START In the Menu – Sterilization screen, press the Start full sterilization button or the Start empty sterilization button, depending on the status of the bioreactor (full or empty).

  • Page 31: Phase 1

    OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 4.5.4 PHASE 1 The rector is now heated up. Phase 1 is completed as soon as the preheat filter temperature is reached. If you stop the sterilization process during...

  • Page 32: Sterilization

    OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 4.5.7 STERILIZATION The reactor is kept at sterilization temperature until the sterilization interval has elapsed: the timer counts down. If during the sterilization interval the reactor temperature drops below sterilization set-point, the countdown timer stops counting.

  • Page 33: Cooling 2

    OPERATOR MANUAL PRESSURE TEST AND STERILIZATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 4.5.9 COOLING 2 After the sterilization interval (during full sterilization) or the natural cooling (during empty sterilization), step Cooling 2 starts. The steam valves are closed, the gas inlet valve is opened, and cooling water is introduced into the reactor jacket.

  • Page 34: Cultivation

    OPERATOR MANUAL CULTIVATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 5 CULTIVATION After the sterilization process is finished, the bioreactor can be prepared for cultivation. Since the contents of the bioreactor must be kept sterile, all actions must be carried out aseptically.
  • Page 35 OPERATOR MANUAL CULTIVATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) Press the button to open the aeration valve. Continue the aeration until DO reading is stable. This will take 15 . . 20 minutes. Press the button to close the aeration valve.

  • Page 36: Connect Addition Fluids

    OPERATOR MANUAL CULTIVATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 5.1.2 CONNECT ADDITION FLUIDS Addition of fluids like acid and base are realized through a Contained Addition Assembly and Push Valves. 5.1.2.1 CONTAINED ADDITION ASSEMBLY The contained addition assembly supports aerosol-free operation and is used for addition of liquids and for inoculation.

  • Page 37: Activate The Control Loops

    Just before starting a new process, the valve is opened (push in the spring knob and push the valve to its opened position). 5.1.3 ACTIVATE THE CONTROL LOOPS Activate all control loops in the ez-Control: Press the Menu button. Select the option Start all controllers.

  • Page 38: Start Cultivation By Adding Inoculum

    OPERATOR MANUAL CULTIVATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 5.2 START CULTIVATION BY ADDING INOCULUM The cultivation is started by adding the inoculum. Like other additions this operation must be carried out aseptically. An easy and safe way of aseptical connection of tube ends is the so-called “Tube Welding”...

  • Page 39: Drain Valve

    OPERATOR MANUAL CULTIVATION ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 5.3 DRAIN VALVE The bottom drain is used for taking relatively large samples from the culture and for harvesting / draining the system after finishing the process.

  • Page 40: Cleaning

    OPERATOR MANUAL CLEANING ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 6 CLEANING Proper cleaning can be considered as a must, in order to avoid problems with the integrity of the system during cultivation. Cleaning before and after processing also gives the opportunity for general inspection of the system and its assemblies.

  • Page 41: Cleaning Auxiliaries And Tubing

    OPERATOR MANUAL CLEANING ez-Control for In Situ sterilizable Bioreactors Project 310420; PS70 IST (AU) 6.3 CLEANING AUXILIARIES AND TUBING Disassemble the bioreactor system and remove all tubing and auxiliaries that are inserted in the reactor (side wall and head plate).

  • Page 42: System Warnings

    In case a power failure occurs that takes longer than the brown out time during operation (running controllers), the ez-Control will detect this event. It will restart with a Power Fail message. The user is logged out (returned to View mode).

  • Page 43: Emergency Stop During Operation

    Log in as System Engineer. The options now enabled. Before pressing the button, make sure that the bioreactor is in a safe condition to restart the process. Pressing the button will turn the ez-Control into an idle mode (no running control loops).

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