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LABFORS 3 OPERATING MANUAL & USER GUIDE

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1. Tm T 14 9 2 M 14 9 3 ON 14 9 4 Se Ier Tero MEER eer ere eee ee eee eee eee ee 14 9 4 1 CS SUIS 14 9 4 2 15 9 4 3 15 9 4 4 APOT NS E 15 9 4 5 15 9 4 6 Elecincal POWE EIE RR 15 9 5 Serial por CORNISH OS ERR En 17 9 6 Memon om 18 9 7 MANGE 19 9 8 gt 19 9 8 1 et ee a te ee 19 9 8 2 ANC 19 9 8 3 Topping up the mechanical 20 9 9 21 9 9 1 t T E 21 9 9 2 en 21 9 9 3 Identification plate and serial 21 10 RENE 22 11 Preparation Sterilization Setup US ic cacssssneassiesanacawcsenstedavarcnonsi
2. 36 Rene 36 12 Switching Parameter on and 37 12 1 Entering or changing Set point alarm 38 13 Sterilization Autoclaving the vessel CtC ccccccccssseecccceeeceeeeeececceeeeeeesceceeeseeseeeeeceeeessauaasceeeeeesssuaaseeeeeeeesaagnsess 39 13 1 Cooling down and replacing the vessel ccccccceeeeeeccceecceeeeeeeeeeeseaeeeeeceeeeeseaeeeeeeeessaueesceeeeeseesesaeaeeseeeeeesaas 39 13 2 Preparation of Corrective reagent lines nennen nnne nnns nnn rrr nns nnn 40 13 9 41 13 4 Polarization electrode calibration amp pO2 42 135 Setup of the Exit Gas Cooler after autoclaving nnn 43 13 6 Setup of the Digital Feed n rrr iS ey mmc ie Lo MOC 44 13 9 Aseptic Sampling 45 13 10 9 46 13 11 Cleaning amp ceeded Uc 47 14 XDDC InstrumemtatiON RE ERU 49 14 4 Main Menu Offline Options ccccccccsseeeeceeeeeeeeeeeeeeeeeessaeeeeeeeeses
3. Selected fermenter 1 Parameter pH Channel 2 HiReading 3544 LoReading 593 Y Dep m Les W 13 14 4 3 PID additional document available for PID explanation Advanced Level 2 3 Explanations 1 Max Value display parameter max measured value 2 Min Value display parameter min measured value 3 Output Max Set the maximum controller output with the value given here as a usually 100 or full range 4 Output Min Set the minimum controller output with the value given here as a usually zero 5 P term Proportional term P The greater the difference between the actual value and the set point the greater the controller output The given value is a multiplier in the differential equation In most applications a suitable P control value is critical 6 Off State The setting here provides a default for when this parameter is switched OFF or when the bioreactor is not in an operational condition 7 Term Integral term 1 Sums the error over time Should the setpoint not be regained using the P term the term enables the output control to carry on working until the setpoint is reached The l term is only set to a small decimal value and small changes can have very large effects 8 Int Limit To ensure that the I term cannot increase indefinitely a limit can be set on the error to be summed Normally leave set at 10096 of full range of
4. 14 5 2 Values This is the default screen when a fermentation is active The screen is described in the Offline Section To quickly reach the Set point option move the cursor to the desired parameter and press ENTER For calibration press the decimal key To switch a parameter OFF go to the Setpoint menu and turn regulation plus Output Active to OFF This will then be reflected in the Parameters screen The parameter is now only measured 14 5 3 Pump As offline 14 5 4 Stop SEE OPPOSITE This allows the bioreactor to be turned off Press 1 then ENTER to end a running fermentation 14 5 5 Diagnose This is the same as Offline Select the required bioreactor 1 5 lt lt Back 4 14 Param Pump Diagnose gt gt Selected fermenter 1 Stirrer 100 2 Off Flow Off Gasmix Off FreeA Off FreeD Off FreeF Click Menu entry If an incorrect choice is made a message screen appears to tell you the selection is not valid Selected fermenter 1 Setpoint Stirrer Off Off pH 2 Off AFoam Flow Off Feed Off Gasmix Off Off Click Menu entry F1 RUNNING lt lt Back Select Values Pump Selected fermenter 1 1 Stop fermentation Turn the rotary L R Scroll Continue F1 RUNNING lt lt Back Select Values Pump 44 Selected fermenter 1 CLOSEDOWN FERMENTER Ent
5. Fl STOP 2 5 Info Select Config Passwrd Receipe 14 1 2 ALL tab Two options are provided in the main screen of the ALL tab They are accessed as usual by clicking navigating to the desired option and clicking Calibrate all pH sensors 9 Calibrate all pO2 sonsors Calibrate all pH sensors Calibrate all pO2 sensors This is an extension of the normal calibration screen which is still available for use with individual vessels as normal Location of the ALL tab The first difference is that as well as entering the reference values each stage of the calibration high low has to be explicitly switched on and will diei automatically revert to off after leaving this screen High Ref Calib Hi Off It can also be seen that the temperature compensation is now provided from Low Ref 4 Calib Low Off just the first vessel s Pt 100 temperature Sensor which can be placed in the P beaker of buffer This is not user adjustable although individual calibration F1 F2 still works in the conventional way Current 4092 4092 Offs 0 048 0 048 Slope 0 0034 0 0034 The display includes information about the status of the probes mv pH for pH and nA reading for pO2 xsl Esc click Exit or value entry Enter When the reading has stabilized individual units must have the calibration Set Reference confirmed by pressing the corresponding bioreactor number Enter each bioreactor number nor
6. 63 I C EE c 63 63 Deseret 63 Jl Dm 63 gt 1 E 63 15 E 64 15 1 Labfors3 Cells Including Gas 00 0000 0000 n nnns nnn 64 15 22 65 15 2 1 Detailed Description of the Infors Gas Mix 66 15 3 Connection to IRIS SOMWANE 67 15 4 Dimensions standard unit T 69 15 5 Dimensions Vessel Top 70 15 6 Labfors and III Minimum Working volume with 72 T 77 156 ROTE 78 15 9 External I O Pin COMMQUMATION x ccccicc ecredecccntecedeereceiciceSeeedsarsedcccaxesbedeisnctdadadaevhcesdeedandachasseaeescae heeedecceedacdeueedscbos 79 9 Installation 9 1 Delivery Information Transport to final location In view of the weight of the complete bioreactor and packing materials it should never be carried by one person alone For moving the
7. Max Value Min Value Output P Terme Error 1 5 Error Error n 1 F1 STOP F2 STOP lt lt Back Setp Calib 38 Selected fermenter 1 Parameter Gasmix Channel 7 Maximum Value Output Max Minimum Value Output Min P Term Off State Zero I Term 1 s I Limit Output Ramp Dead Band Step Size D Term s Neg Factor Eval Time s Turn the rotary L R Scroll Continue Tuning a PID loop a simple approach 1 Make changes one step at a time Changing the proportional P term will exert the most influence on the process variable Increasing the P band too much will cause a gradual drifting of the actual value and too small a band will cause hunting e g for pH a little acid is added followed by a little alkali followed by acid 2 The integral band should normally be a small value eg and changed a little at a time 3 The D term is often set to zero to begin with and may not be needed Alternatively where large changes take place rapidly a high value may be necessary 4 An Evaluation time can be set which determines the time interval between recalculation of the PID terms 5 Integral limit can sometimes be increased if control is not good 6 The Negative factor can introduce a bias eg for pH a negative factor of 10 would imply a strong base was being used and 0 1 that a strong acid was being used 7 A bas
8. Membranes are fitted to ports used for piercing with a needle eg Inoculation Sample device and reagent bottles are connected to vessel if they will be autoclaved together 2 Lines to sparger sample pipe and reagent inlets are all clamped SHUT pH pO2 electrodes are capped or covered with foil Drive shaft top is covered with foil cap SAMPLE DEVICE Filter fitted onto needle for pressure equalisation Inlet needle connected to harvest pipe in vessel via silicone tubing which is clamped SHUT Spare bottles prepared REAGENT BOTTLES Or Filter fitted onto needle for pressure equalisation Dip connected to inlet line in vessel via silicone tubing which is clamped SHUT Line is prepared for aseptic coupling to the vessel following autoclaving and is clamped SHUT Bottles are filled with reagent at the correct strength and clearly labelled The same principles apply for medium feed and harvest bottles for continuous culture although these are almost always sterilized separately INFORS NT 13 2 Preparation of Corrective reagent lines If the reagent bottles have been autoclaved with the vessel they should be connected to the inlet pipes on the top plate The stages for setting up the pumps to deliver liquid reagents acid base and antifoam are as follows 1 Unwrap tubing from bottle and unclamp 2 Pass tubing FROM bottle to the slot at the underside of the correct pump labelled Acid Base AF and Fe
9. LABFORS 3 OPERATING MANUAL amp USER GUIDE 43 INFORS EQUIPMENT Read this FIRST AN SAFETY COMPLIANCE SPECIFICATIONS MAINTENANCE amp SERVICING 1 E ow t B of 79 Th Operating Manual Labfors 3 e 1 CE conformity declaration INFORS equipments are CE labelled from the factory and supplied with an EC declaration of conformity INFORS equipment complies with the following Directives amp Standards Directive Machinery 89 37EC Directive Electromagnetic compatibility 89 336 EEC IEC61326 ti e AME Directive Low Voltage 2006 95 EC EN61010 2 Quality Management Systems All INFORS equipment is manufactured in accordance with INFORS quality management system which is certified by BVQI in accordance with the requirements of ISO 9001 3 _ Testing All INFORS equipment undergoes electro mechanical operational testing before it leaves the factory The exact nature of the tests varies according to the equipment type All equipment is delivered with a signed test certificate The tests described are conducted in accordance with the procedures set out in INFORS quality management system and in accordance with international classification companies 4 Applicability This is specified in the specifications table included within this section of the document If any of these specific sections appears to be missing for your equipment please contact INFO
10. 00E iri 8 of 79 d Le ic Operating Manual Labfors 3 e Double click on the IRIS icon from a desktop shortcut the Programs section of the START menu or the Infors AG folder An opening screen will then load and IRIS will open with the default user logged in A first only the Alarms window and User Log windows open within the main IRIS window TE mnt l FERMEHTER The main window comprises a menu bar with a toolbar beneath The bottom oy Fani o part of the window is a status display with descriptive information about menu options and details of the active fermentation The position and range mde dea of options provided on both the menu and toolbar areas is fully Windows 2222 compliant See the IRIS Operating Manual for further information ira a Wu FE n E uem 15 4 Dimensions standard unit 7 0 17 1 T gel ame a standard unit E A Dimensions Labfors 3 Pa a a CH 4103 Boallmngen 341 0561 425 77 00 T2926 ww infarg H com Al rigo m particular the cognmghid in th amp document beng 10 OR SAG Boomingen Swit eran dy Tre document giento you solely or your personaluse according i fie agreements iors concluded with you every use whith exceed the abo ned age ihe transmission ip hird parties he making of copies photocopies exiracts
11. at the bioreactor operating panel see description down right or via the software IRIS e With biologically dangerous organisms the disposal or subsequent use of the organisms must comply with laboratory safety procedures e Empty the corrective reagent lines means direction through manually expelling the liquid from each pump in turn acid base antifoam feed Turn off the device at the main switch e Removing the cultivated organisms in a living and uncontaminated condition for further experiments requires that you connect a harvest pump to the sampling system outlet and pump the cultivation fluid into a sterile container over e Remove the tubing of the corrective reagent bottles without dismantling it from the pump head 2 only using gloves and safety glasses and rinse the tubing thoroughly with water Before further dismantling the containers they should be sterilized in an autoclave according to local biological security directions before cleaning page 46 of 79 Push Clamp Change bottle amp replace m AL 4 To stop an active bioreactor go to the Stop tab F1 RUNNING F2 STOP lt lt Back Select Values Pump Selected fermenter 1 1 Stop fermentation Turn the rotary L R Scroll Conti nue Then press 1 and confirm with Enter instructions on the screen F1 RUNNING F2 STOP lt lt Back Select Values Pump Selected fermenter 1 CLOSEDOWN FE RMENTER Enter Click
12. block to the desired parameter Use the number keys for data entry or the key to toggle between On and Off 14 1 4 Password Pin The password Pin code allows to specify different authorisations for different users Critical parts of the control system can be protected from unauthorised access There are 5 different password levels see manufacturer s pin setting Level 1 2 3 and 7 have each an individual and modifiable pin code password which is possible to activate New pins can only be activated and defined in the offline mode no active process OFF It is possible to change a level during an active process Simply choose an option that needs a more advanced level Once entered a pin code is only valid until the Auto Log Out Time has expired this is manually adjustable Afterwards the system falls back automatically to level 0 Log OFF and all functions are locked again As soon there is a function that needs a higher level a new log in is necessary It is also possible to login to any level after Auto Log OFF during an active process Entering a false pin code leads to an automatic Log Out A pin code consists of a maximum of 9 digits The numbers are shown as on the display Below the first level whose pin is activated no pin is needed The pin code of the next higher and activated level must be entered when higher levels are deactivated OFF and the lower ones are protected This setting is not recommended because
13. key confirms your input and immediately leaves the menu option 3 Usethe ESC to leave the menu option and any changes including those values confirmed previously are not accepted 4 Help on How to proceed can be found on the bottom edge of the If communications have been correctly established a diagnostic red light will flash rapidly above the green mains light to indicate this fact If the LED flashes slowly the communication display 4 se 5 Tabs on the top line of the display give main options gt gt forward or lt lt etween the operating panel and the back base unit is not properly established 5 The main part of the screen is accessed by clicking the rotary knob which can then be used to move up and down options 6 Some screens allow direct entry of a numbered option from the tab 8 Click the knob again to go back to the tabs or press the Esc key 15 02 2008 Operating Manual Labfors 3 e page 31 of 79 11 4 1 Starting a fermentation To start the bioreactor select an individual bioreactor bioreactor 1 4 if necessary then move the rotary switch clockwise and the next tab to the right Start is highlighted Turn the rotary knob to the Param parameter field Click to enter the menu Choose each parameter by turning the rotary knob Enter the Set points by using the numeric keys Use the key to turn on their output ON Esc Enter or click to leave the menu After starting the biorea
14. 00 FreeH Turn the rotary L R Scroll Continue Click Menu entry Type in setpoint Value screen F1 STOP F2 STOP F1 RUNNING F2 STOP lt lt Back 4 Calib PID PConf Name SCasc lt lt Back Select Pump Stop Diagnose Selected fermenter 1 Selected fermenter 1 Parameter pH Channel 2 Value Setpoint Setpoint 7 Alarm Lim 5 Stirrer Temp compensation OFF 20 C 2 AFoam Outp active Off Flow Feed Gasmix Turn the rotary L R Scroll Continue Click Menu entry INFORS 11 Preparation Sterilization Setup and Use MAIN COMPONENTS 1 LCD Display 11 Status lights incl Mains power ON 21 Water inlet manual control 2 Keypad 12 Inlet amp Rotameter assembly 22 Exit gas cooler manual valve 3 R8232 port 13 OPTIONAL mass flow valve atc 23 Main power socket amp 4 Knob 14 Motor connection socket 24 Stainless steel drip tray on base 5 Connection to Base Unit 145 15 Analag comms connections rear 25 Drive Motor on top plate Memory Card slot 16 OPTIONAL analogue pump 26 Port fittings 7 Digital Perietaltic pumps 17 Water Inlet 27 Mechanical seal on drive shaft Manual Button for pump motor 18 Water inlet for jacket nr cold finger 28 9 Cables 100 pH pOs Foam 18 Water retum for jacketed vessels 29 Baffles 10 Operational Amplifser Module 20 Water inlet Exit Gas Cooler 30 Sparng
15. 35 44 LoReading 593 0 0033 0 0094 Current Value Slp mV pH 57 2 Ref Temp 20 F1 STOP F2 STOP lt lt Back PConf Name SCasc Selected fermenter 1 Parameter pH Channel 2 HiReading 35 44 1 593 0 0033 0 0094 Current Value Slp mV pH 57 2 Ref Temp 20 Click Menu entry F1 STOP F2 STOP lt lt Back Setp PConf Name SCasc Selected fermenter 1 Parameter pH Channel 2 HiReading 35 44 LoReading 593 20 0033 0 0094 Current Value Slp mV pH 57 2 Ref Temp 20 7 Click Menu entry INFORS 11 5 2 Dissolved Oxygen 2 Electrode preparation e Before first use and after every longer unused period the base of the electrode must be unscrewed to refill the electrolyte into cartridge pressed onto inner glass section depending on type of electrode e Remove cartridge by turning bottom section over and GENTLY pressing metal mesh in base Fill silicone cartridge with electrolyte provided remove cover from under bottle cap first and make sure air bubbles remain gt Electrolyte is vitriolic e Press onto glass section gently some liquid will come out and reassemble e For storage gt 4 weeks empty the electrolyte and store dry 11 6 Reagent bottle preparation The sterilization of supply vessels takes place in an autoclave As a rule borosilicate glas
16. Electrodes are described in the manufacturer s literature in the box with the electrodes There you will find information about cleaning and regeneration The pO electrode membrane cartridge MUST be filled on first use then checked and refilled with electrolyte after every 2 3 sterilizations for normal operation 9 8 2 Chemical resistance The housing was tested with various substances In connection with the following substances no significant changes were found Water Inorganic acids phosphoric nitric sulphuric 1096 solutions Organic acids formic vinegar and lactic 1096 solutions inorg alkali sodium hydroxide 1096 solution inorg Chemicals in aqueous solution Hydrogen peroxide 1096 Calcium chloride and concentrated bleach hydrocarbons benzene Benzene motor oil amp diesel The housing should not be in contact with the following substances for prolonged periods of time Acids alkalis and inorganic chemi cals in higher concentrations than given in section Methanol Ethanol Acetone Cyclohexanol Ethyl acetate Ethylene glycol acetate Methylene chloride Perchloro ethylene Carbon tetrachloride INFORS MT 1 Look for liquid in the tubing 9 8 3 Topping up the mechanical seal 1 The seal is lubricated with glycerine and must NEVER be allowed to run dry 2 Some liquid in a loop of silicone tubing at the base of the drive hub on the top plate shows if any glycerine 1 still present
17. INFORS A Please ensure a Risk Assessment is carried out according to your safety regulations before using the equipment A Live steam under pressure can cause severe burns and physical damage due to release of pressure suddenly 6 2 Safety points relating to installation amp use A Electrical connections should only be installed and fitted by a qualified electrician to current electrical safety regulations A Installation of all services lines should be made only with pressure resistant tubing retained with suitable tubing clamps A Authorization for use of an oxygen supply and its operation in accordance with your own safety guidelines are the responsibility of the customer A If previous operation was not subject to either an unexpected loss of power or closed down by turning off individual parameters but switched off using the main ON OFF switch then restarting the unit is analogous to a return of power after a failure and the last set points will be used for control This means that the stirrer can operate and possibly that the acid and base pumps may deliver reagent Therefore a fermentation must always be terminated by explicitly turning off individual parameters A Never use the main ON OFF switch to end operation Never remove a top mounted motor or work on the drive shaft of bottom drive units with the mains switch set to on A Always use gloves when removing a top mounted motor after a period of use as it is likely
18. Loosen both grub screws on the stirrers with the provide Allen wrench do not unscrew to eliminate biological organisms in the space between stirrer and stirrer shaft in order to clean the shaft and stirrer blades separately Check all O rings and replace them if necessary 2Do not clean the top plate in a washing machine as water must not get into the bearings For severe problems the drive hub should be sealed against liquid ingress to allow automated washing 2Disconnect all electrical connections with the console before wiping or cleaning the surface of it Reconnect them only after the console is completely dry again The console must not be doused with water Specifically check the insulation of the antifoam tube and replace it if necessary 2 Be particularly carefully when sliding the tube into the clamp nozzle If the bioreactor is not used for a long period 1 month or more then store the individual parts separately Strictly follow all manufacturer s instructions concerning operation cleaning and storage of the sensors NOTES 14 XDDC Instrumentation 14 4 Menu Offline Options Rotate to L or R Abort key or back to tabs The X DDC panel is mounted above the main instrument cabinet It can S Es Numbers be removed completely An RS485 connector links the panel to the base unit via a cable with a pair of RJ45 connectors An RS232 port is available under the bottom left of the panel for programming
19. Pump Diagnose gt gt 14 3 1 Closedown from here requires moving to the Stop tab and pressing 1 followed by eee ENTER to confirm closedown 2 Flow Gasmix FreeA FreeD FreeF 14 3 2 Parameter Freel The Parameter tab is really a gateway to the series of screens which provide the opportunity to alter a large number of options relating to one particular parameter eg temperature or pH F1 STOP F2 STOP ion j j i Back Calib PID When the fermentation is active this screen is not necessary as access to lt lt Back individual parameters be gained via the Values screen Therefore the Selected fermenter 1 option is not displayed when the bioreactor is in Running mode Setpoint 500 The main part of the screen is accessed by clicking the rotary knob A Alarm Lim 1200 cursor block now appears by the first parameter on the list and rotating the knob to the right will take the cursor block to each parameter in turn Select the chosen parameter by clicking when the cursor is next to it Pressing Esc will take you back to the tabs Selecting lt lt Back will return you to either the main menu options bioreactor inactive or the Values screen if active Turn the rotary L R Scroll Continue 1 5 F2 STOP A new set of tabs now appears with the screen for the leftmost tab lt lt 15 FID Setpoint display
20. by a plug and socket should be installed and fitted by a qualified electrician to current electrical safety regulations Installation of water and air lines should be made only with pressure resistant tubing retained with tubing clamps jubilee clips INFORS LABFORS Tubing set up guide OPTION Recirculating chiller Must have pressure relief amp bypass if not an open bah Exit Gas design Cooler Jubille clip l Rapid coupling i of base connector To Drain s EE Overflow or lt Connect to 4 cold fir finger base unit Vater m circulator Double IN gt s jacketed Eg vessels only H 1 mm 15 02 2008 Operating Manual Labfors 3 e page 17 of 79 9 5 Serial port connections The X DDC system can communicate directly over the serial connection or with a computer running the IRIS software RS 232 On the underside of the instrument panel on the left side a male 9 pin connector is provided Do NOT use the serial port connector in the base unit section this is reserved Pin Configuration Pin 2 receive data RD Pin 3 send data TD Pin 5 Signal ground 5232 Connections 1e 2 30 3e ce 7e 9e 5e Description PC Side PC Side Labfors 25 9 pins 9 pins Ground 7 5 5 Tx Rx 3 2 3 Rx Tx 2 3 2 Tx Rx INFORS MT 9 6 Memory Card The memory card is located in a slot under a cover at the left s
21. end to the corresponding serial port of your PC laptop or desktop Screw in tightly 3 An earth lead may be provided on the cable For desktop PC s This can be connected to a casing screw For laptop computers the casing is plastic so this is not possible Install the software as for any Windows 98 NT3 5 2000 Me amp XP PC based program INFORS HT 4 Insert the IRIS CD If autorun is set for the PC it should start automatically 5 If not locate the file Setup exe on the CD and double click to run 6 Follow the instructions given choosing standard installation and accepting the defaults for the simplest installation 7 Place a shortcut to the IRIS icon on the desktop for easy access GS N 7 The software installs in approximately 27MB of hard disk space but additional capacity must be available for the data files which will be created It is not possible to predict exactly how much space is needed as different conditions of use will apply to different fermentations As an example data logging 5 parameters for one hour at one minute intervals each a relatively high log rate for most parameters will create a file approximately 50K in size A simple control sequence will create a file of only a kilobyte or so All IRIS data files have the structure FILENAME iri and show the Infors IRIS icon IRIS creates a new file for every fermentation It is advisable to use a different directory than C programmes for installation
22. gloves safety goggles and a face mask as appropriate to the degree of microbiological risk A The nature of the microbiological and chemical risks associated with the use of individual units cannot be assessed by the manufacturer and its specification is the responsibility of the user A The environmental hazards associated with the use of individual units cannot be assessed by the manufacturer and its specification is the responsibility of the user 6 3 Safety points related to testing the equipment A Shoxic the previous fermentation not have been stopped using the Intervention Stop menu but by turning off at the main power switch analogous to a power failure the set points in force at the moment of termination will be used on re start This means that the stirrer can run A The main power switch should not be used as a functional ON OFF Never put your hand into an operating unit risk of injury due to high rotational forces A No reagent lines to be left connected to the pumps if the pump operates or the contents spill out there is risk of damage from acid or base A Handle glass vessels reagent bottles and other glass components with care to minimise the risk of breakage or other damage resulting is sharp edges A Do not apply excessive pressure when handling any glass components in case of breakage and consequent sharp edges A Ensure hair loose clothing etc cannot come into contact with any rotating parts 7 E
23. many functions would ask for the Administrator pin 7 A high level pin combination leads to free access to all functions that are equal to or below the chosen level Certain aspects of configurataion can only be made when in Service mode only accessible on Administrator Level 7 Please note The number of people with supervisor access Administrator has to be limited Anyone with this level of access must not make changes unless they are certain of the outcome If there is any doubt they MUST first consult an INFORS representative or the INFORS Service department compliance with this point could render your warranty void 1 5 F2 STOP Info Select 11 4 Passwrd Receipe Language 0 GB 1 DE 2 FR 0 Tube empty time in s 0 Auto Backlight Off Buzzer enable On Accept remote On Parameter Alarn Off Adjust contrast Click Turn the rotary L R Scroll Continue Password screen eg Level 3 Fl STOP F2 STOP Info Select All Config Receipe Activ Change Off Off Off Off On Off Off 2 3 Advanced 7 Supervisor Current Level 22 Auto Log Off time 5 60 Turn the rotary L R Scroll Continue The following information is shown on the display when no sequence 1 active manufacturer s setting Pin Active Change Pin Level 1 On Off On Off Level 2 On Off On Off Level 3 On Off On Off Level 7 On On Off Auto Log Off time s 900 Current level 0 no functions unlocked c
24. rapid coupling connector Water inlet to double jacket rapid coupling connector Place a pipe from the overflow to the drain using the silicone tube 10mm inner diameter INFORS MNT Single walled vessels should arrive fitted with a heater pad Connect the optional cold finger in the top plate with the rapid coupling connector with the base unit Control valve exif gas cooler Connegtion to 5 Connect the rapid coupling connector to the water inlet on the base unit lower right side 5 Connect the outlet of the exit gas cooler to a flexible pipe and run this to the drain sink or return to a thermocirculator if used INFORS If not already fitted locate and fit the heater pad around the vessel Connect the heater pad plug to the socket on the lower left INFORS NT 4 Connect the motor to the base unit using the steel DIN type socket at the middle right of the base unit front one 5 Mount the motor on top of the vessel turning until it locates on the pin in the hub Locating pin on drive 6 Plug in the mains cable and switch on the electrical supply 7 Connect the rotameter outlet pipe above glass scale to the sparger tube with holes in the vessel with flexible tubing and turn the rotameter needle valve knob anti clockwise until gas can be seen bubbling though the liquid Filters are not needed at this stage but the inlet filter ma
25. to be HOT A Always remove the tubing from the peristaltic pumps when cleaning the base unit or vessel in case acid or base may be accidentally released Warning do not autoclave reagent lines filled with corrective agent in case the tubing bursts Use water instead As part of the cleaning regime the corrective reagent tubing MUST be rinsed with water Storage and autoclaving of tubing containing acid or base represents a hazard to the user and can damage the tubing A Take care regarding overpressure following autoclaving Tubing can burst or come off inlet pipes Ensure a pressure equalisation line is present for each reagent bottle capped with a small filter A Take to avoid stick injuries when handling the piercing needles for the inoculation and reagent inlet lines plus fitting the antifoam electrode AN When inoculating or adding any peripheral by piercing a membrane ensure the flame does not come in contact with hands electrical wiring etc in the vicinity of the vessel top plate A Always work to GMP and observe other appropriate standards Observe all safety issues relating to hazardous chemicals biological material and equipment under pressure especially points regarding skin and eye contact A The equipment is only to be operated by suitably qualified and trained personnel both in terms of equipment use amp microbiological expertise A In normal use operators should ware appropriate safety clothing
26. will select the process value or option next to the cursor if a further screen can be reached Fi STOP F2 STOP IMS Select All Config Passwrd Receipe The bottom 2 lines of the display list options of what to do next and how to access the choices INFORS AG Copyright c Aug 07 2007 INFORS XDDC SYSTEM LABFORS 3 14 1 1 Initial Choices from the X DDC Menus offline eee Info Information about the X DDC version of the firmware in your VERSION 1 51 bioreactor IS also provided here Labfors 3 Controller Copyright Infors AG Version 1 40 Nov 09 2006 Select This is the first screen where options are available to select an individual bioreactor multi bioreactor system A number of offline Fut HER SERO options are listed as a series of tabs running from left to right across the screen Moving the rotary know to the right will highlight the name of each in turn and clicking the knob downwards where appropriate will allow the main part of that screen to be accessed Ferm2 Ferm3 Ferm4 Active bioreactors show the current actual values and inactive units display a line of dashes Arrows indicate that more options are available Back indicates that the original options can be reached again by turning the rotary switch left Other tabs available are All Config Password amp Recipe Turn the rotary L R Scroll Continue of 79 ING B Sm 15 02 2008 Operating Manual Labfors 3 e
27. 216 ASZZ 005 ASZZ 005 only ASZZ 005 cooler D 12 D 12 D 19 D 19 D 19 Oo Z o m o 2 I gt c i O w O O 2 N e e Clamping device 21500 aa Labfors Il and Minimum Working volume with electrodes ie 15 6 NW 200 15 02 2008 Operating Manual Labfors 3 e page 73 of 79 NW115 1 2 2L EE 11 Clamping device 31145 L 225 220 SETEFFF SSS TO Oberflache X CH 4 03Eolmiagen OO 41 7151425 77 09 sam E NG 007589 SASEA l 4 237 at Ped Ne tion yeti 2 a2 M 164516421 hee ere ce wmm ame Dem T7 ici videor gt 9149 1 Lio 57 RELSE AS page 74 of 79 Manual Labfors 3 e c _ 3 pop E eornm 1 15 r 2 D rre 2 Se Oy 1 d lt lt C gt NW115 2 3 3 6L Zi 22 M x 5222 s g
28. 3 To replenish the glycerine no glycerine is visible anymore in tubing before autoclaving pull off the tubing from the longer arm 4 Connect a syringe filled with glycerine to the silicone tubing and pump glycerine in until it can be seen emerging from the open long pipe 5 Remove the syringe and remake the loop 2 Fill with glycerine if necessary 3 Replace tubing 9 9 General Points Components not supplied directly by INFORS All components sourced from outside the INFORS group of companies have been tested before incorporation into our products As a general rule INFORS only uses proven components In exceptional cases it is still possible that faults can occur in other manufacturer s components In such cases INFORS cannot accept any liability other than our normal warranty obligations We ask our customers for understanding and do of course always strive for the highest possible quality This exclusion does not apply to safety matters 9 9 1 Reselling For reasons of operator reliability legal considerations smooth and continued supply of spare parts and the protection of INFORS intellectual property rights the customer is obliged to inform INFORS of any change of ownership or re sale In return for this service by our customers INFORS offers to support the new user and supply training for the new operator s INFORS reserve the right to refuse to supply maintenance support or spare parts if this duty i
29. O electrode can be used it must be connected to the base unit and the electrical power turned on for a minimum of 6 hours Without this polarization time any attempt at calibration will be unsuccessful Calibrationa Zero Before fitting e With special gel from the manufacturers METTLER INGOLD The electrode tip is pushed into the gel and the zero point set using the Low Reference calibration This calibration is guaranteed by the manufacturers for applications where the partial pressure of oxygen Is gt 5 Where the same electrode is always used a single calibration is enough unless a problem occurs e With nitrogen the electrode must be placed in a measuring cylinder and introduced Or b After fitting By gassing with Nitrogen in the growth medium This is achieved after the vessel has been cooled to the operating temperature and the reagent lines are closed Calibration of the 100 point follows on from this 100 Calibration see description opposite As arough approximation calibration of 100 in air prior to autoclaving is possible especially for cell culture work The oxygen partial pressure value as opposed to the zero point claibration depends critically upon the values for Temperature the higher the temperature the lower the O2 solubility Stirrer soeed Air flow rate Pressure the higher the pressure the bigger the O2 solubility These parameters must be set to the correct operational values
30. Push until metal to metal contact can be heard and felt to ensure accurate readings pH sensor 4 Connect the cable by pulling back on red locking ring on the cable and pushing connector down onto the electrode 5 Screw into place Buckling or twisting the cable while mounting will destroy its delicate sheathing and lead to faulty measurements pO 6 Connect cable by aligning orange line in connector cap with orange line on top of electrode connection 7 Twist locking ring on cable down and clockwise to connect to electrode Removal is the reverse gt Buckling or twisting the cable while mounting will destroy its delicate sheathing and lead to faulty measurements Important do not touch the metal ends of the banana connectors with your bare hands without grounding yourself first by touching the equipment Antifoam 8 After autoclaving fit the RED banana connector to the hole in the top of the electrode and the black one into the small hole in the top plate made to accept the connector 9 The electrode is hollow so a silicone tubing line for antifoam can be fitted not acid or base as these conduct 2Important Always loosen firstly the clamping device before adjusting height of the sensor damaged sensor insulation would cause faulty measurement and lead to permanent addition of antifoam medium 13 4 Polarization electrode calibration amp pO2 control Polarization Before the dissolved oxygen p
31. RS and this can be rectified Please be aware that if the equipment is acquired second hand from an original user it may have been modified upgraded and enhanced such that some details of the configuration may differ to those described in this manual We will provide any help and information necessary to bring the documentation up to date but individual options may not be the standard ones supplied by Infors In this case it is the responsibility of the previous owner to supply any additional manuals configuration information and safety related items INFORS disclaims responsibility for all equipment that is not in original condition i e modified by the user without prior agreement from INFORS 15 02 2008 Operating Manual Labfors 3 e page 4 of 79 5 Identification plate _ 27 _ The identification plate is located on the housing of every piece of equipment The identification plate must never be removed from the equipment If the name plate is removed it is not possible to identify the equipment and it will not be possible for warnings contained in this manual to relate to the specific applications for which the equipment is used The serial number of the equipment is also displayed on the vessel identification plate for stainless steel pressure vessels 5 1 Vessel identification plate 6 Safety Points 6 1 Ge
32. Specially for use with eg E coli bacteria The minimum value can be higher than maximum value for reducing the feed rate by pO2 control E g Min limit 100 max limit 0 Exit gas filter To drain Water inlet coupling Example The desired flow rate is eg 0 2L Hr 3 Therefore the pump must run for approximately 37 of the total time Desired rate Max rate x 100 A set point of 37 leads to 22 2 seconds of dosing time in every 60 seconds Example The actual value displayed for the fermentation is pH 6 5 The external measured value is 6 9 Therefore the difference is 0 4 In the Field Offset the displayed value relates to the actual value of 6 5 It must therefore be corrected by adding 0 4 to the offset to give a new value 6 9 INFORS In Use and Afterwards What you should check before inoculation 138 Inoculation e Correct Temperature e Correct Stirrer soeed cascaded to oxygen control e Correct pH Correct pO partial pressure e Have you checked the growth medium concentration is correct after autoclaving losses not Cells version e Are any heat labile supplements to be added eg glucose solution e Are the pumps primed Before inoculation the correct operational status should be verified Are the corrective bottles connected and is there enough reagent to last for the desired fermentation time The precise method of inoculatio
33. Stop the fermentation Esc Return to overwiew screen Esc click Exit or velue entry Enter 13 11 Cleaning amp Storage After autoclaving the vessels the leftover liquid can be disposed down the drainage Remains of dried foam and proteins become denaturated and sticky Fill up the vessel with 0 1 N NaOH and stir it for about 2 hours 2 Attention Strictly follow the laboratory safety procedures while working with NaOH If foam has reached the gas cooler Remove the gas cooler from the top plate and put it then for 4 hrs into 0 1 N NaOH Rinse it afterwards thoroughly with water and put it then into a ultrasonic bath for about 2 to 5 minutes Finally wash through with 7096 Ethanol and rinse again with water The tubes must be completely empty no corrective reagent should remain before cleaning and rinsing the tubes and pump heads Basically all screw in components of the top plate should be dismantled and cleaned 2 Do not forget to loosen the clamp nozzles before you remove the sensors Any use of force can destroy the sensors especially the insulation sheath of the antifoam sensor which has to be handled very carefully After removing the retaining nuts take the top plate off the vessel at right angles and put it on a clean underlay Inserted parts of the stirring system must not come off 2 Attention Any deformation of the stirrer shaft can damage the mechanical seal and will harm the bearings in the hub
34. Time s 1 The desired setpoint value can now be entered conforming to the range defined in PID Pumps can have a setpoint range of eg 0 100 and the steps can be set in 0 1 increments However an analogue pump cannot turn less than 1 rom below this a fault condition will occur For example if the analogue pump can operate from 0 100 rom the minimum possible step will be 1 The integral analogue AMSP 300 can operate from 0 100 rpm using Bioprene tubing of ID 3 2mm with a wall thickness of 1 6mm This has a flow rate of 48ml min at 60rpm range approximately 1ml min at 1rpm to 80ml min at 100 rpm The same general procedue would apply to any signal having a 0 20mA input or output eg input from a biomass probe dissolved CO2 analyser glucose probe exit gas analyser etc NB Parameter names can be changed by an Infors specialist if desired Output ml h 22 000 48 600 102 800 156 400 210 000 266 600 323 200 374 200 425 200 479 500 933 800 ml min 0 367 0 810 1 713 2 607 3 500 4 443 5 387 6 237 7 087 7 992 8 897 Digital Pump other tubing sizes ID 100 flow rate output mm 0 63 1 14 1 42 1 52 2 05 Material Silicone Silicone Silicone Marprene Marprene ml h ml min 32 4 0 54 88 1 47 154 6 2 58 180 4 3 01 322 8 5 38 14 5 Online Options 14 5 1 Start This is described in a former section When online it displays the current status of the parameters eg setpoint or OFF
35. a low set point eg 4 The gas should flow for up to 24 seconds in every minute and the acid pump will be seen to turn 6 Attach the gas inlet pipe to the inlet air filter and the outlet for CO2 if fitted to the filter on the pipe needle for head space gassing 7 Set Gas mix as the first cascade level in the setpoint menu for pO2 Adjust flow rate to 0 1 0 3 vessel volumes per minute Ensure the pO2 electrode has polarized properly minimum 2 hours before turning pO2 control to ON for the actual fermentation F1 STOP F2 STOP lt lt Back 5 1 PID Selected fermenter 1 Parameter Gasmix Channel 7 Setpoint 0 Alarm Lim 100 Outp active Off Turn the rotary L R Scroll Continue F1 STOP F2 STOP lt lt Back 135 Calib PID Selected fermenter 1 Parameter p02 Channel 3 Setpoint 80 Alarm Lim 100 Outp active Off Cascade Channel l Level Off 1 Stirr O 2 Level Off 5 Flow 0 3 Level Off 7 Gasmi 100 100 Turn the rotary L R Scroll Continue INFORS MT 15 3 Connection to IRIS Software The LABFORS system can communicate directly over the serial connection with a computer running IRIS software The following communications protocol is used Communication settings BAUDE RATE 9600 DATA BITS 8 STOP BITS 1 PARITY BIT NONE FLOW CONTROL NONE 1 Connect one end of the serial cable to the RS232 serial port at the bojtom of the operating panel left side Screw in tightly 2 Connect the other
36. all setpoints entered ready for operation the next task is to sterilize the vessel and peripheral items by autoclaving Inactive Active 13 Sterilization Autoclaving the vessel etc The Standard Labfors3 bioreactor is sterilized in an autoclave at 121 C for a minimum of 30 minutes and typically for one hour For water circulation units the vessel jacket must be filled for heat transfer and efficient sterilization During this process approximately 10 of the liquid volume will be lost due to vaporization This must be allowed for either by over diluting or adding extra liquid during inoculation Reagent boitles the sampling device and any other peripheral items are sterilized in the same way generally along with the vessel Tubing should NOT be filled with liquid Any heat labile components are normally filter sterilized and added afterwards by injection or with the inoculum If the whole medium is heat labile several strategies are possible Sterilize the vessel with a small quantity of PBS phosphate buffered saline enough to cover the electrode tips and remove it after autoclaving using a sample harvest pipe Add medium at the correct strength amp correct volume e Ifthe autoclave is too small and the vessel must be tilted it is possible to put a very small amount of liquid in the vessel and allow that to generate a moist environment to protect the electrode tips Either of these methods could be used for ste
37. and data transfer A slot on the left side houses a memory card which is used to store all the or setup and calibration data etc The system must NEVER be used without Glick knob a memory card in place Switching on the mains power at the base unit to accept also activates the rotary DDC panel The bioreactor should be inactive at from tabs or AUTO this point A POST Power On Self Test screen appears briefly and all to main screen the fields should show PASS next to them If not please switch Off On and back confirm number again after checking a memory card is fitted entry or accept Screen Display of Rotary XDDC active F1 RUNNING The top line shows the tabs which lead to screens with more options lt lt Back Select ZETA Pump Stop Diagnose e The main section of the display is accessed by clicking the rotary knob inwards Other than the Values screen which shows live data of all the 2 oim process parameters there are options to be set using the numeric keys or the key to toggle options such as On Off or Auto matic e Return to the tabbed area is by pressing the Escape key or clicking the pu Ore rotary knob depending on the screen options po2 Off e Navigation up and down in the main section of the screen is by moving fior i rotary knob left up right down As the screen scrolls a cursor Feed Off block indicates the current position Gasmix Off e Clicking using the rotary knob
38. as well as ihe storage an data camies af amy nature requires the prar wrtren consent by INFORS AG 2 m Ta oD c H 5 i un 5 e ue ea X a Y 5 5 t t 2 L 5 u T T c zi T gt i T i a xm INFORS MT 15 5 Dimensions Vessel Top plates Sparger Sample Sparger Sample Spare Dz212 Pg 13 5 8 Foam gas _ Probe cooler 4s probe Inoculation e cooler needle C 12113 3P3 15 t Sparger 0 10 2 Pt 100 Inoc needle Sample Base Pt 100 pO2 452 04 4x Antifoam 12854 4 en Dz12 Pg13 5 Bx D 12 P913 5 6x Feed Exit gas cooler Sparger NW 150 NORMAL NW 150 MAGNETIC PA qm iy i Bn EIE ems 1a LT 1 d t C gt 15 02 2008 Operating Manual Labfors 3 e page 71 of 79 Inoculation needle 2 Sample Ini spare Pt 100 gt needle i S e a Exit gas cooler Spare Antifoam probe inlet probe Acid Alkali e Exit gas par inlet cooler Acid Alkali NW 200 NORMAL NW 200 MAGNETIC NW 115mm NW 150mm NW 200mm diameter Vessel Tot V AMKL 2005 2 L AMKL 206 3 6 L AMKL 214 3 6 L AMKL 216 7 5 L AMKL 218 13 L m 2 Option 3 2 Option 3 2 Option 3 Ratio h d for 1 8 for TV 3 2 for TV 1 6 for TV 3 1 for TV 2 1 for TV TV Exhaust gas ASZZ 216 ASZZ
39. ation ensuring the needle section is covered with a sheath or foil or bag It should be connected to the bottle vessel to be used for transferring the inoculum 11 8 3 Antifoam Electrode This is packaged separately and should arrive assembled ready to fit onto the top plate It detects foam on the surface of the culture using conductivity and this can trigger a pump to deliver a dose of liquid antifoam reagent 1 Fit electrode into 19mm or 12mm small vessel magnetic drive port complete 2 Adjust height by loosening top part of clamp and pulling electrode gently upwards Take care not to damage the transparent sheathing around the electrode clamp still too tight is better to be too low pull up as the converse is not possible due to the contamination risk 4 After autoclaving fit the RED banana connector to the hole in the top of the electrode and the black one into the small hole in the top plate made to accept the connector 5 The electrode is hollow so a silicone tubing line for antifoam can be fitted not acid or base as these conduct 11 8 4 pH 4 Place electrode into 13 5Pg port fitting and screw into place 5 Ifa longer electrode is used add a clamping adaptor and adjust the height to suit the vessel size before screwing adaptor just tight enough to hold the electrode securely Do NOT use tools 6 Cap electrode with red cap for autoclaving 7 Connect the cable by pulling back on red locking rin
40. atly when all services outlets including the necessary pressure reduction system and appropriate filters where necessary are already installed by the customer 9 4 Connections 9 4 1 Gas Supplies For cell culture systems or bacterial systems with oxygen supplementation the appropriate gas supplies must be available at the time of installation and commissioning gt The gas supply must be pressure regulated max 2 0 bar and have suitable rotameter if constant gas flow into the bioreactor is to be main tained Unless specified with the bioreactor supply of these components is by the customer Authorization for use of an oxygen supply and its operation in accordance with your own safety guidelines are the responsibility of the customer Barbed pipes for push fit connection to services are provided in a separate You may need bag Push these into the corresponding sockets in the base unit Flexible tubing for these pipes is provided as part of the Service Kit 9 4 2 Water Inlet Cold water supplied at a pressure of gt 1 5 bar enters from the left side of the base unit of the bioreactor Push on connector pipe 10mm secured with a jubilee clip gt Only cooling water with no more than a slight hardness can be used exclusively The use of drinking water supply for cooling purposes is possible as long as it is very soft or soft Never use demineralised or distelled water Contact your INFORS Service for questions
41. before the calibration maximum values for this specific fermentation to provide the 100 calibration point Control POz2 control can be mediated by Stirrer speed Standard Mass flow control valve altering air flow rate Supplementation with oxygen pulsed from a valve Gas mix with CO O and air Cells version Control of feed rate special setup required A Cascade function exists as a special section in the pO2 Parameter Setpoint menu This can be up to three stages providing sequential cascade with the ability to define which parameters is to be used at each stage by its channel code Each parameter used can have a starting set point defined within selectable minimum and maximum limits Individual steps can be turned on or off in Sequence depending on need The order of listing of the parameters determines the sequence of their usage Switching the first parameter in the cascade off deactivates all the other stages No parameter can appear in the list twice EVEN if it is switched off Calibration of the 2 Electrode 100 First you must A 2 Setpoint 80 Outp active Off Cascade l Level Off 1 2 Level Off 5 3 Level Off 7 Locate the drive motor on the top plate Set the stirrer speed RPM set point value to the maximum to be used during the fermentation Switch on air supply pump if necessary Set the air supply on the rotameter to the maximum value to be used eg 1 5vvm Set temp
42. concerning water quality and calcification 9 4 3 Water Drain A Thermocirculator for supply of chilled The drain outlet is on the top of the bioreactor jacket if applicable It water at eg approximately 10 C requires a clear fall to the drain sink with no back pressure For single NB Must have a pressure relief and walled vessels with a heater pad a cold Finger may be fitted bypass system fitted Push fit onto glass the overflow connection arms 12mm or the drain arm of cold finger as appropriate 9 4 4 Air gas Inlet The air inlet can be found on the right rear side of the bioreactor Use only gt clean dry oil and dust free air with a pressure of gt 0 7 bar 2 Push on Connector pipe 6mm Secure with a jubilee clip So m EE 2 Never connect the gas supply in any way which bypasses the pressure reduction valves Secure gas tubes with hose clamps pua 1 L 9 4 5 Exit Air A diaphragm type air pump for oil free The exit air leaves via the disposable filter attached to the exit gas cooler if supply of air eg 0 15L min fitted If oxygen supplementation is used take care to pipe the exit air away safely to avoid explosion risk 9 4 6 Electrical Power 230V 50Hz 10A single phase supply for standard LABFORS or 115V option The power supply should be clean and constant if not it is recommended you fit suitable filters and or a UPS to the mains power supply Electrical connection
43. cording to the demands of the pH controller Alternative options and arrangements are available so please check the delivered configuration matches your requirements eg the mass flow valve has the right range Note In the gassing system for O2 and CO2 needle valves have been integrated after the magnetic pulse valve On delivery the valve is open It can be adjusted manually by turning the screw clockwise to restrict the flow Setting Up 1 Connect the gases to the corresponding inlet pipes and secure FIRMLY with jubilee clips to ensure there are no leaks expense and safety reasons Make sure the metal pipes on the inlet have enough exposed metal to connect firmly when pushed to fit 2 Connect the outlet pipes for the mixed gas and CO2 but do NOT connect to the vessel just yet 3 Start the bioreactor with a test set up using only pO2 control at first and set Flow to ON providing a suitable set point eg 0 5NL min in the Pump Valve menu so gas can be felt escaping from the outlet pipe 4 Test each gas in turn by adjusting to get Air O2 and N2 to flow individually When each gas flows adjust the pressure roughly with the regulator on the gas cylinder or compressor Each gas needs an input pressure regulated to 0 7 0 9 bar Make sure inlet pressure does not exceed the max of 2 0 bar Gasmix Setpoint 100 Nitrogen only flows 0 Air only flows 100 Oxygen only flows 5 CO2 is set by turning pH control to ON and giving
44. ctor by pressing Enter the active screen is now displayed showing information about each parameters actual value setpoint and controller output a number from either 0 100 or 100 to 100 relating to how hard the controller must work to bring the actual value to the set point value Stopping fermentation Closedown bioreactor bioreactor 2 Emergency use the main ON OFF switch being aware of any hazards making this difficult Alternatively use the switch at mains power socket Normal closedown Go to the Stop tab and press 1 All parameter functions stop Press Enter to confirm when dialogue screen appears bioreactor closes down and offline menu options are shown Fl STOP F2 STOP RINT 111 Config Passwrd Receipe Fermi Temp 24 0 Stirrer 180 Selected fermenter 1 24 0 Stirrer 7 0 po2 0 00 Flow off Gasmix 0 00 0 00 FreeD F1 RUNNING F2 STOP lt lt Back Select Pump Stop Diagnose Selected fermenter 1 Value Setpoint Output Off 1 Off Selected fermenter 1 1 Stop fermentation Turn the rotary L R Scroll Conti nue F1 RUNNING F2 STOP lt lt Back Select Values Pump Selected fermenter 1 CLOSEDOWN FE RMENTER Enter Click Stop the fermentation Esc Return to overwiew screen Esc click Exit or velue entry Enter INFORS MT 11 5 Before Filling 11 5 1 Preparat
45. d will help 13 5Pg ring amp port Full details of vessel sizes proposed port allocations and lengths of appropriate electrodes can be found in appendix Assuming the objective is to begin a new fermentation a basic preliminary is the cleaning of the vessel and its peripherals Check the fit and connections of individual components before and after cleaning to prevent damage The vessel top plate is removed by undoing the four nuts around the circumference of the vessel and carefully lifting the top plate up far enough to clear the glass section The sealing of the vessel openings is achieved using O rings These are usually located in the parts to be installed such as the port fittings electrodes etc Only with 13 5Pg ports you will find them in the recess of the plate Please ensure that before installation these O rings are undamaged To make fitting and release easier wet them with alcohol The following parts can be removed before cleaning Baffles compress top slightly and pull out of vessel glass Sparger connected to the top plate Stirrer shaft connected to top plate with impellors e Port fittings temperature sensor also sample valve device can easily be disconnected from the top plate NB The drive shaft can be removed by taking the top plate from the vessel and resting it on the bench with the drive shaft uppermost Use a spanner to turn the flattened section of the drive shaft whilst k
46. e during a 9 second cycle eg 50 4 5 seconds dose 4 5 secs delay With the parameter Feed 1 A value between 0 100 ON time is given With the parameter Gasmix This controls either oxygen supplementation for bacterial systems or the switching of Air Oxygen Nitrogen for the 3 gas mix system used for the Cell culture systems The set point is this case is the percentage oxygen in the gasmix Set to 100 only Nitrogen would flow Setto 0 only Air would flow Set to 10096 only oxygen would flow Normally this is set automatically by the dissolved oxygen control loop Remote control IRIS If setpoints are sent remotely from the IRIS software the external values will overwrite anything entered at the control panel of the bioreactor Please bear this in mind when attempting to alter set points Values Active Inactive CLE Esc Accept Inactive Active 1 5 F2 STOP lt lt Back 5 Calib PID Selected fermenter 1 Parameter p02 Channel 3 Setpoint 80 Alarm Lim 100 Outp active Off Channel Min Max Start Limit Limit StPoint Cascade l Level Off 1 2 Level Off 5 3 Level Off 7 Turn the rotary L R Scroll Continue Fl STOP 2 5 lt 11 PID Selected fermenter 1 Parameter AFoam Channel 4 Dose time Wait timr Regulation On OFF Manual IRIS Outp active Off Turn the rotary L R Scroll Continue 14 4 2 Calibration pH Value Exp
47. ed Other options are Calib rate PID and Pdiag Taht tti Parameter p02 Channel 3 The options available in the Parameters submenu are described in detail in XN point 80 Alarm Lim a separate section Outp active Off Cascade Channel Which Parameters are available l Level Off 1 Stirr O Pre allocated 2 Level Off 5 Flow 0 3 Level Off 7 Gasmi 100 100 e Speed e Temperature e PH e pO2 PO2 is Special e Antifoam A cascade option at the bottom of the e Feed Setpoint menu allows a range of Flow option parameters to be used sequentially for e Gas Mix control of dissolved oxygen at a given 4 Free for eg setpoint weight The numbers before parameter name e Redox refer to the channel code of the e Analogue Output for Pumps parameter to be used for this control at Biomass Optical density a particular level This must be entered manually 7 of 79 Lye 14 33 Pump This screen shows the total on times in seconds for active pumps Click to enter the main part of the screen A cursor block will appear by the first parameter option Acid and Base pumps are displayed separately The running time for the pump s associated with that parameter will be displayed Only the Feed pump can be set manually to deliver manually using a does and delay cycle set in the Feed parameter Explanations 1 Pump 1 Acid pump Pump 2 Alkali pump 2 Antifoam pump T
48. ed so the white connector stops it from pulling though 3 Pull on white knob at bottom of pump head and then lift the black cover right upwards Pass the Marprene yellowish tubing around the pump rollers Replace black cover and re clip Pass the tubing through the slot in the top of the black cover so that the white connector again prevents the tubing from pulling right through 7 Press the black knob with power switched ON and the pump motor starts manually Keep pressing the button and push the black cover backwards so the pump rollers rotate This primes the pump When liquid from the pump falls into the vessel release the button and black cover The pump is ready for use Su The pumps are labelled acid base etc Make sure the correct reagent bottle is attached to its appropriate pump Should any liquid base acid etc leak into the pump housing remove the tubing instantly and clean the pump head After removing the crosshead screw rear right side of housing you can extract the steel drip tray for cleaning 2IMPORTANT Tubing used in the pump heads must have a wall thickness no greater than 0 8mm for correct operation INFORS 13 3 Connection of electrode cables Pt 100 temperature sensor 1 This is the only sensor on Labfors3 bioreactor which needs no connector 2 Itis added to the vessel after autoclaving by pushing the end of the electrode into the special 10mm pocket on the top plate 3
49. eeeeeeeeeeesseeesseeeeeeessseessesseeeeeeesssaeaaeeeeeeeessaas 49 14 1 1 Initial Choices from the X DDC Menus 49 Mie Eo 50 ir Ai ec E 51 EE 51 14 1 5 Recipes advanced level 3 5 14 2 Selected Bioreactor 55 55 14 3 Values Screen Online cccccccceeeseeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeesseaeeseeeeeeeeeesesseaeeeeeeeeessaas 56 56 WES TAN RR TETTE 56 co RUND P 57 14 3 4 Diagnose Advanced Level 2 3 58 D MEN se c 59 sae ey 59 1442 Calibration 60 Ir VP d 61 additional document available for PID 99 8 9 9 61 14 4 4 Additlonal Parameter SelUD 24 4 eia e 62 ir Mm
50. eeping the bottom section still The mechanical seal is exchanged by releasing the spring locking ring using a small hexagon key and taking the various seal elements way When replacing it is critical that you DO NOT touch the seal faces or clean them with acetone to remove any grease from fingerprints etc If you are able to make this exchange yourself we can provide information which provides step by step instructions on request Alternatively this exchange can be made as part of a preventive maintenance contract of 79 W AT 2 15 02 2008 Operating Manual Labfors 3 e 11 4 Switching ON at the base unit and Display The operating panel is removable from the support frame at the back of the Before turning on please note all safety instructions first base unit Undo the single bolt at the The main power switch on the front right side of the instrument housing back of the panel to release It can press down to switch ON A green light immediately shows in the small now be placed on the bench window above the switch on the base unit if power is present f not check the socket and fuses The unit automatically performs a self test diagnostic on start up If everything is satisfactory PASS will appear next to the tested components such as etc If one or more components show FAIL switch off at the mains switch wait a few seconds and then switch on again to re star
51. elp to facilitate work and save time calibrating sensors This special feature of XDDC is described in section ALL tab Rinse the electrode again and keep it protected and in electrolyte until ready to place it in the vessel no times subject the electrode tip to damage while calibrating or rinsing NB Slope is pH units per mV and so has a small value F1 STOP F2 STOP lt lt Back Start Gain Pump Diagnose Selected fermenter 1 Stirrer 2 Flow Gasmix FreeA FreeD FreeF FreeH 2 Click Menu entry 4 Click Menu entry 5 Click Menu entry Menu entry 1 5 F2 STOP lt lt Back Setp PConf Name SCasc Selected fermenter 1 Parameter pH Channel 2 27 HiReading 35 44 2 LoReading 593 0 0033 0 0094 Current Value Slp mV pH 57 2 Ref Temp 20 F1 STOP F2 STOP lt lt Back Setp PConf Name SCasc Selected fermenter 1 Parameter pH Channel 2 High Ref HiReading 3544 12 LoReading 593 0 0033 0 0094 Current Value Slp mV pH 57 2 Ref Temp 20 Click Menu entry F1 STOP F2 STOP lt lt Back Setp PConf Name SCasc Selected fermenter 1 Parameter pH Channel 2 HiReading 20 50 LoReading 593 Current Value Slp mV pH 57 2 Ref Temp 20 1 5 F2 STOP lt lt Back Setp PConf Name SCasc Selected fermenter 1 Parameter pH Channel 2 HiReading
52. eparations for Autoclaving The vessel is cleaned and assembled Medium is added to the required working volume possibly with a 1096 over dilution to allow for losses during autoclaving The top plate fittings are added to the assembled vessel Assemble all sensors etc needed before autoclaving to reduce risk of contamination 11 8 1 Exit gas Cooler This can be found packaged separately from the vessel It prevents excessive loss of liquid from the culture through evaporation The inner section has a series of cooling late to aid condensation of water which is returned to the vessel 1 Depending on the vessel size and type this can be fitted directly in either a 12mm or 19mm port 2 Insert the base of the cooler into the base and push into place Screw into the port fitting 3 Adda short length of reinforced silicone tubing to the topmost pipe and fit the exit gas filter green tape towards the pipe Secure with cable ties at both ends 4 21115 VITAL the exit gas line is left open CLAMPED shut as this allows pressure equalisation following autoclaving gt explosion risk 11 8 2 Inoculation Port This will be packaged separately along with a bag of membranes of a Suitable size 1 Remove port closure and insert a membrane of the correct size 12 or 19mm 2 Screw collar into place to hold membrane securely Re fit port closure into collar for autoclaving 4 Prepare the inoculation needle for steriliz
53. er 111 Checking Services Connections For OPERATION it is assumed that a correct installation and operational tests have been carried out and also that there are no mechanical alterations to the unit Please make sure that you always inform us if the bioreactor is to be used for unusual applications eg higher temperatures pressure control etc gt Take care that The supply of air amp water are guaranteed and they are of sufficient quality Check the pressure regulation is correct 11 2 Safety Points gt If the current fermentation is not shut down from the start stop menu i e by simply using the main ON OFF switch switching ON again will be analogous to a power failure and the operating conditions which existed before the interruption eg setpoint values will be used immediately on re start That implies that the stirrer and reagent pumps eg acid amp alkali will also run Therefore gt Never use the main ON OFF switch to end operation When setting up or dismantling after a fermentation Never use the mains switch as an operational ON OFF Never handle the motor system with mains power ON Only pick the motor up by the handle or by using gloves after prolonged use the casing will be HOT heater pad if used may get hot to the touch if elevated temperatures are use take care when removing No reagent lines to be left connected to the pumps it may be that there is still some acid or alkali residue i
54. er Click Stop the fermentation Esc Return to overwiew screen Esc click Exit or velue entry Enter f C ur Ul 4 of 79 15 02 2008 Operating Manual Labfors 3 e 15 APPENDICES 15 1 Labfors3 Cells Including Gas Mix The special version of Labfors3 for culture of mammalian and insect cells has a number of differences to the standard unit Not all of these may be present for each installation No baffles for low shear Magnetic coupling system Infors axial system Slow speed option max 20 300rpm Spin filter for cell free removal of culture supernatant Single marine type impellor several designs for low shear and good mixing at slow speeds Sinter sparger or special system for smaller air bubbles Dissolved oxygen control by air flow control using a thermal mass flow control valve e Special mixing options for dissolved oxygen and pH a Atwo gas supplementation system for oxygen and carbon dioxide b Athree gas mix unit for air oxygen and nitrogen A three gas mix unit with an additional Open axial magnetic drive gas inlet for carbon dioxide These options can be used with or without mass flow control Some or all of these supplementary gases may be added to the head space through a separate inlet Most options are self explanatory but the magnetic drive and spin filter need some additional explanation Magnetic Coupling E This k
55. erature and pressure if necessary Wait 5 10 minutes for the medium to saturate with oxygen Go to the calibration display press for a one point calibration and set the value at 100 Selected fermenter 1 Parameter pO2 Channel 3 Alarm Lim 100 Channel Min Stirr 0 Flow 0 Gasmi 100 100 Turn the rotary L R Scroll Continue Cascade Setup in pO Setpoint Screen Max Start Limit Limit St point Param Min Name Cascade 1 Level OFF 0 Stirrer 100 1000 500 2 Level OFF 5 Flow 2 10 5 Level OFF 7 100 100 0 13 5 This is used to prevent excessive losses of liquid from the culture in the form of water vapour carried out in the exit gas stream The Infors thermodynamic cooler has a special internal structure which increases the efficiency of the re condensation process Setup of the Exit Gas Cooler after autoclaving The cooler is connected by braided flexible tubing from its lower inlet pipe to the specific water supply pipe in the base unit A manual valve controls the flow of cooling water When the outer wall of the condensor feels cold to the touch the flow rate is adequate A second pipe of normal silicone tubing links the outlet at the top of the condensor to a drain sink This must have an unobstructed fall to drain at a shallow angle 2 Only cooling water with no more than a slight hardness can be used exclusively see details in appendix 13 6 This is the forth pump additi
56. eters are switched on click the rotary knob to accept and start the fermentation The setpoints and status for the parameters is set in the Param screen see below lt lt Back 494 Pump Diagnose Selected fermenter 1 Temp Off Stirrer Off Off po2 off AFoam Off Flow Off Feed Off Gasmix Off FreeA Off FreeB Off FreeC Off FreeD Off Click Menu entry Switching parameters on amp starting the fermentation H Repeat for other parameters Accept amp start fermentation Values Shows parameter current actual value current set point Controller output as a 100 to 100 typically 121 Entering or changing set point amp alarm limits F1 STOP F2 STOP lt lt Back Start Pump Diagnose gt gt Move to the Param tab when the bioreactor is inactive and click to enter the main part of the screen which shows a list of parameters Not all of Selected fermenter 1 these may be available for use on individual systems according to specification 2 Gasmix FreeA FreeD Move the cursor using the rotary know to select the desired parameter eg and click to enter the parameter setup display with Setpoint as the first option Different parameters have different options esp pH and antifoam eg see opposite Click Menu entry Click to enter the screen and the cursor will move to the first field for alterat
57. f the screen then use the rotary knob to navigate to pH and click Move to the Calib tab using the rotary knob and click to enter the main screen 2 The cursor will automatically move to the High Ref field Use the numeric keys to enter the value of the high reference buffer eg pH 7 3 The electrode should now be placed in a beaker containing this buffer solution Unless the Pt 100 temperature sensor 1 also placed in the buffer AND pH temperature compensation is switched ON the buffer temperature is assumed to be 20 C Press Enter and the cursor moves automatically to the High Reading field A number is shown which is in the range of 0 4095 and represents the digitised signal from the pH electrode When this reading is steady press Enter to accept the value 4 Move the cursor to the Low Ref Field 5 Rinse the electrode using a wash bottle with distilled water and place in the lower value buffer solution eg 4 Repeat the steps as for the High value buffer and after the final Enter to accept the Low Reading value 6 move the cursor past the slope and offset fields rotary switch two positions to the right and click to accept the calibration 7 The Esc key aborts the process and leaves the original values in place A one point calibration only uses the High Ref and High Reading values Move the cursor past the Low Ref and Low Reading fields without altering them and click to accept gt Using the ALL tab will h
58. for fungi etc Option 2 allows retrieval of a setting from a particular location on the memory card Options 3 requires a file name to be given and a simple terminal program to be available on the PC not Hyperlink A pattern on the display indicates data transfer Option 4 allows a file name to be selected for upload to the memory card A pattern on the display indicates data transfer 2 Options to send data to and receive data from recipe files stored on a PC may only be used when a computer is connected and NO bioreactors are active Otherwise errors in operation can occur due to a system crash Data cannot be read into a card when the bioreactor is active Option 5 Explanations 1 The number of timeouts within a fixed time interval when a request for communication has not been answered 2 Not Ackn Is the number of time the recipient has not understood the request for communication 3 Bad rec is the number of time a communication packet has not been fully understood an indication of a bad contact or cabling fault 4 Pack Rec is the total number of communication packets successfully received 5 Pack Sent is the total number of communications packets successfully sent 6 With Output it is possible to switch between data going IRIS software or to a serial printer 7 Comm Speed shows the serial baud rate and should remain set at 9600 this does not influence performance 8 Parity Opt shows the parity opt
59. for use when making enquiries 8 CONTACT DETAILS Infors AG Rittergasse 27 4103 Bottmingen Switzerland Tel 4161 425 77 00 Fax 41 61 425 77 01 eMail headoffice infors ht com Internet http www infors ht com Infors UK Ltd Unit H Dovers Farm Lonesome Lane Reigate RH2 7QT Tel 44 1737 223100 Fax 44 1737 247213 eMail Infors uk infors ht com Internet http www infors ht com Infors GmbH Tannenweg 2 D 85254 Einsbach Tel 49 8135 83 33 Fax 49 8135 83 20 eMail infors de infors ht com Internet http www infors ht com Infors SARL 6 rue Marcel Paul F 91742 Massy cedex Tel 31 169 30 95 04 Fax 31 1 69 30 95 05 eMail _infors fr infor ht com Internet http www infors ht com Infors Italia S r l Via Prestinari 2 20158 Milano Tel 39 0239 31 02 13 Fax 39 0239 31 72 04 Mail infors it infors ht com Internet http www infors ht com Infors Benelux Logistiekweg 20 02 NL 7007 CJ Doetinchem P O Box 613 NL 7000 AP Doetinchem Tel 31 314 364 450 Fax 31 314 378 076 E mail infors oni infors ht com Internet http www infors ht com m d ks Contact details of our local dealers world wide can be found on our web site INFORS MT HOW THIS MANUAL IS ARRANGED e Thefirst section deals with installation requirements next section tells you how to bring the LABFORS into operation in a step by step manner including e The third section c
60. g on the cable and pushing connector down onto the electrode Screw into place 11 8 5 1 electrode into 13 5Pg port fitting and crew into place 2 Ifa longer electrode is used add a clamping adaptor and adjust the height to suit the vessel size before screwing adaptor just tight enough to hold the electrode securely 3 Cap electrode with aluminium foil for autoclaving 4 Connect cable by aligning orange line in connector cap with orange line on top of electrode connection 5 Twist locking ring on cable down and clockwise to connect to electrode Removal is the reverse 11 8 6 Air Inlet 1 Fita 0 22 micron filter red line on filter to the top of the sparger pipe using reinforced tubing secured at both ends with cable ties Note the side of the filter marked INLET should be on the top facing away from the vessel 2 The air input filter may only be connected loose fitting with the tube that is connected to the rotameter 2 Danger of overpressure 3 For autoclaving this inlet line MUST be clamped shut with eg a gate clip 15 02 2008 Operating Manual Labfors 3 e page 37 of 79 12 Switching Parameter on and off Move to the Start tab when the bioreactor is inactive The screen now shows a list of parameters with Off after the name Use the rotary knob to move to those parameters you wish to switch ON and use the key to toggle the setpoint On Off for that parameter When all desired param
61. his is under the control of the antifoam probe It is turned on in the Pdiag display for the AF parameter 3 Medium dosing pump The flow rate is set via the Set point Menu Feed or from an external signal It must be explicitly turned on in the Pdiag display Calibration Any pump can be calibrated to display weight in grams or millilitres delivered by first selecting the pump then pressing the decimal key A new screen can then be seen Enter a pump speed in percent 0 100 eg 50 Enter a pump rotation eg 500 for the pump eg a value of 500 will allow delivery of approx 2ml using standard tubing sizes based on Labfors3 pumps When Calibration is turned from OFF to ON using the key the message RUNNING appears in the display After the calibration run time has expired end enter the weight dispensed or ml delivered and the pump factor is automatically worked out to allow display in the desired units The example opposite causes the calibration to last for a total of 1 minute as 30 seconds is 50 On If a pump has been successfully calibrated the value displayed is shown with ml after the numbers This indicates a pump factor which is not equal to one If a pump is uncalibrated s for seconds will appear after the name to indicate this i e pump factor 21 F1 STOP lt lt Back Select 1 5 Diagnose Selected fermenter 1 duration volume Displayed volume depends on calibratio
62. ic manual approach to tuning a PID loop when you don t have any previous settings as a guide relies on setting the P value so the proportional band is as wide as possible and the and D terms are zero 8 Gradually narrow the P band until the controller causes oscillation in the process value Now adjust the P term back a little and repeat the process for the term starting with a long time interval and gradually shortening it until oscillation again sets in If required the D term can be added Digital pump Marprene tubing D2 97 x 4 39mm wall 0 8mm part 22899 14 4 4 Additional Parameter Setup Feed Pumps The Feed parameter can be used to set the operation of the digital pump option The tubing used can be varied but MUST have a wall thickness of 0 8mm and a Shore hardness of 64 Flow rates for the standard Marprene tubing are shown opposite with some information on other sizes If the optional analogue pump is used by allocating one of the free channels this is what should be considered e Control range This defines the range of control signals to be used for the controller output eg 0 and 100 corresponds with a control output of 0 20mA e Volume If the flow rate of the pump is known this can be entered directly This is normally in ml min In the PID screen these values should be used Maximum Value 100 Minimum Value 0 P Term 1 D Term s 0 Term 1 s 0 Limit 100 Dead Band 0 Neg Factor 1 Eval
63. ide of the X DDC operating panel Its function is to store configuration data about the fermentation eg calibration values amp PID settings etc It does NOT store experimental data Sufficient memory is available for the card to hold the configuration of multiple bioreactors if the system is used in this way The facility is provided for the data on the card to be stored on a PC and uploaded when required see Section Recipe The battery inside the card buffers the data and has to be exchanged regularly durability up to 2 years Otherwise data stored on the card will be lost whenever the bioreactor is switched off Save your data see section 5 1 before exchanging the battery Push the button to release the memory card 2IMPORTANT After taking out the old battery immediately replace it with the new one 1 Remove the card from the slot in the X DDC panel housing noting the orientation of the card A firm pull may be needed to release the card 2 Your bioreactor MUST ALWAYS be turned off before removing the memory card 2 Put your fingernail or eg a thin penknife blade into the small crack above the area containing the screw hole and pull backwards Pull out the memory card battery holder is revealed with the battery sitting in position Note the orientation of the battery and exchanged for a new one Touch a new battery only sideways when exchanging 4 the memory Ensure the w
64. ind of magnetic drive is based on experience of the development T S of the multi bioreactor SIXFORS Up to date technology provides a secure strong coupling in a small space The INFORS axial magnetic drive is crevice free due to its open design and therefore can be cleaned in place Spin Filter A spin filter option is available for the removal of culture supernatant free of cells This is important for applications such as the production of monoclonal antibodies where the cells remain in the culture vessel and the culture medium containing the antibody is removed The filter part is made of a mesh with a pore size too small for animal cells to pass through It is mounted on the drive shaft using the small screw in the base collar at a height which ensures it dips into the culture The rotation of the drive shaft creates a centrifugal force which keeps the cells from settling and growing on the mesh A separate harvest tube is mounted in the top plate so that the end is inside the cup created by the mesh filter Culture supernatant collects in this cup free of cells and can be collected via the harvest tube Fresh medium can be added via either the Labfors3 in built feed pump or external pump INFORS MT 15 2 Setting up the Gas Mix Options a Control of dissolved oxygen by mass flow control This is the simplest system for cell cultures and may be sufficient for cells with a lo
65. ion which is the setpoint value lt lt Back PID Selected fermenter 1 The value can now be changed using the numeric keys and then Enter is Parameter pH Channel 2 used to confirm the new value Use the rotary switch to move the cursor to other fields such as alarm limits A symmetrical alarm limit can be set PNE here in the units of the parameter eg for pH with a setpoint of 7 an alarm limit of 1 would be 1 ph unit l e between pH 6 8 and there would be no Temp compensation OFF 20 C On alarm Outp active Off For control as well as measurement of any parameter ensure that Output Active is also switched to ON Exception regulation for stirrer speed must be switched off OFF Click to accept and the use the Back tab to return to the main offline menu tabs Inactive The Programming of additional parameters is analogous the procedure described for pH above A point to consider concerns the stirrer speed and gas flow set point values used when working with media which are prone to form foam Set the values appropriately i e not too high Determine the highest rate necessary following sterilization by increments whilst using antifoam reagent NB When the bioreactor is running the parameter can be selected from the Values display by clicking to enter the main part of the screen moving the cursor to the required parameter and clicking This leads to the parameter setup sub menu tabs With
66. ion and calibration of pH electrode The pH electrode must be calibrated before insertion into the vessel before the vessel is filled at the latest If the pH electrode is to be used for the first time the silicone seal over the diaphragm is removed using the small craft knife included with the electrode shaped like a key This can be scraped off rather vigorously to reveal the small white dot of the diaphragm beneath The silicone rubber sheath is removed from the bottom of the electrode and the red cap at the top replaced by the red connector attached to the cable on the base unit The electrode must be calibrated before sterilization preferably using a two point High Low regime The electrode is connected to the cable at the left of the base unit which has the red co axial connector see diagram opposite for location of the right cable Hold the cable on its black collar Push carefully the connector onto the electrode Turn the red connector 2 3 turns until the cable is connected firmly with the electrode Then start with the calibration Mounting the pH Electrode in the Vessel Fit the calibrated electrode directly into one of the 13 5mm Pg ports or use an adaptor if necessary for longer electrodes SNOTE Hand tighten ONLY do NOT use tools The instrumentation is switched on and following initialisation the Param tab is selected 1 followed by the pH parameter Click to enter the main part o
67. ion for serial communication and is set to NO as standard 9 Interval gives the time between data strings being sent to the printer in minutes Min 1 Minute Does not work with IRIS 10 Last Param determines the number of parameters sent to the printer Does not work with IRIS 11 With Alarm you can decide if alarm messages will be sent to the printer Does not work with IRIS 12 Unit ID here you can assign a unique ID number to the bioreactor When more than one bioreactor is connected to the PC by a common serial port all bioreactors must have a different Unit ID beginning with 0 and matching the bioreactor list within IRIS 13 Bad rec IRIS is the number of time a communication packet has not been fully understood an indication of a bad contact or cabling fault 14 Pack Rec IRIS is the total number of communication packets successfully received Option 6 Display screen to show how many bioreactor base units are present and their current status 1 working if switched on and functioning normally F1 STOP F2 STOP Info Select All Config Passwrd 255582 Output IRIS Comm speed 9600 Parity Opt No Timeouts 5 Interval 1 Not Ackn 3 Last Param 10 Bad rec 12 Alarm Off Pack rec 1271 Unit ID 0 Pack send 1274 Bad rec 0 Pack Rec 0 Turn the rotary L R Scroll Continue 1 5 F2 STOP Info Select All Config Passwrd 5072382 Working St Working St Absent Absent Esc click Exit or value entry En
68. ional lifetime No operations beyond those expressly stated in this guide are authorised by Infors as being suitable for the equipment Infors will not be held responsible for any equipment which has been improperly used maintained modified or repaired nor for any consequential losses arising Additionally users are responsible for ensuring that the equipment is used in accordance with safety procedures applicable to their work and that the bioreactor is free of any biological or chemical contamination if an examination by Infors staff is requested All the housing covers of the base unit and control panel are as they may cover critical areas only to be removed by personnel who have been explicitly authorised by INFORS to do so Individual safety warnings are given throughout this manual where appropriate and clearly labelled with the following symbol D These points cover both potential risks to the operator and the machine by mishandling or incorrect use 2 Please ensure a Risk Assessment is carried out according to your safety regulations before using the equipment If in doubt on any aspect of the use of this equipment or its suitability for an application please contact your local subsidiary or distributor Service and technical information can be obtained by contacting INFORS AG See our web site www Infors ht com for latest contact details Please note the serial number of your unit s plus the approximate date of purchase
69. lanations 1 High Ref Calibration buffer 1 e g 7 0 2 Low Ref Calibration buffer 2 e g 4 1 3 Slope as pH units mV hence the small value 4 Offset or Zero point Using this value a drifting pH electrode can be rehabilitated 5 Current Value This shows the current pH value as a digital number converted by the transmitter 0 1024 mid point pH7 512 6 Slp Slope of the electrode in millivolts per pH unit 7 Channel This is the channel No in the controller card assigned to pH This cannot be altered from this menu 8 Hi Reading The pH value shown as a digital reading 9 Lo Reading Ditto 10 Ref Temp Temperature of the buffer A one point reading only uses the High Ref values This can simply be whatever the current reading 1 although accuracy across the range may then be compromised Move the cursor over the Low Ref and Low reading without changing the values The calibration for pH and dissolved oxygen 100 should be done for every fermentation Temperature readings from a Pt 100 are much less likely to need recalibration Temperature compensation for pH can be switched On or Off If Off a buffer temperature of 20 C is assumed If On the Pt 100 temperature sensor should be removed from its pocket in the vessel and placed in the buffer with the pH electrode Parameters such a Feed with no sensor to provide feedback clearly need no calibration Fl STOP F2 5TOFP lt lt Back
70. lating to a fermentation process can be stored on the Memory Card You can store the data from the card onto an external computer via RS232 either for security or when you wish to run another sort of fermentation with out an additional card for exchange 1 Store a X DDC configuration on a memory card Up to 9 configurations can be stored on a single memory 1 9 with 0 as the default which cannot be used All setpoints calibration data PID settings etc are stored 2 Retrieve the configuration from a memory card to a X DDC system Be sure to safe current values before reloading an existing configuration back into use After using this option switch the bioreactor Off and On again to ensure the settings are re initialised 3 Send the contents of the memory Card to a computer 4 Read the contents of an archived Memory Card from a computer back into the card in the bioreactor 5 Comms Statistic and Settings lets you set up communications protocol for a two way transfer of data between rotary DDC and a PC running IRIS software or a one way transfer to a printer and view the status of this connection 6 Controller card options For each Labfors3 base unit this should display the card number and Working if the bioreactor is switched on Option 1 allows a configuration to be stored directly onto a memory card Up to 9 complete configurations can be stored for retrieval as needed eg one profile for bacteria one for yeast one
71. mally in the order 1 to 4 when the reading is stable Info Select All Config Passwrd Receipe High Ref 9 Calib Hi On Low Ref 4 Calib Low ATTENTION Ref Temp from F1 23 9 1 Current 4092 Offs 0 048 Slope 0 0034 mV pH 57 2 Esc click Exit or value entry Enter Switch ON calibration mode 14 1 3 Configuration The Configuration menu is where a number of housekeeping utilities are found 1 Language settings 0 English 1 German 2 French 2 The Auto backlight is switched on approx 20 minutes after the last key 15 pressed it switches off until a key is once pressed 3 Buzzer enable is the acoustic alarm depending on the alarm settings of the equipment independent on the software IRIS by selecting OFF the acoustic alarm is deactivated 4 Accept Remote When connected to a computer either data only is sent OFF or control commands can also be accepted ON 5 Parameter Alarm if set to ON any parameter which goes into an alarm state will trigger the external alarm relay built into the 25 pin D socket of the base unit If OFF only generic alarms such as a power failure will be output externally Also an alarm in software IRIS will be shown via this output 6 Adjust contrast Click to enter the screen This option can be set visually using the UP DOWN keys to increase and decrease the contract Click to access the main screen Use the rotary knob to move the cursor
72. meter accuracy 4 96 in Pressure 1 2 bar absolute ange P 1 2 20 C depending on rotation speed ER 2 15 8 Utility Requirements INFORS HT Labfors 3 Utility requirements to Labfors 3 fermentor Cell amp Bacterial T Air in process air culture gazing NS 6mm 1 87 Connection Male adapter or 10 C 20 C 10 C 20 C 10 C 20 C 10 C 20 C Pressure fbar no backpressure ____ Male adapter or Its on top of the Fermenter jacket it requires a clear fall tothe drain Gasing for Cell fermenter 15 25 1 5 2 5 1 5 2 5 15 25 15 25 CO2 air N2 O2 Male adapter or NS 6mm 1 8 NS 6mm 1 8 NS 6mm 1 8 NS 6mm 1 8 NS 6mm 1 8 Recommended maximum soft water up to 1 5 CaCO3 INF ORS 15 9 External I O Pin configuration Top view of Labfors 3 Connector Digital Out open collector mox V mA Analog Out 0 20mA 13 Alarm Com 25 Alarm O 12 Alarm 2 Analog 0 20mA 0 5 0 10 24 Digital Out B s Qu gt 4 10 Analog In FreeA 4 9 Analog In FreeB 5 4 8 Analog In FreeC 6 lt 1 7 Analog FreeD 7 8 ell 6 Analog Out A rT el 5 Analog Out B T TAE ONN 24 4 Analog Out C alog In FreeA 4 9 3 Analog Analog FreeB 5 155 2 Analog Out E 14 GND 4 Analog Out F Analog I
73. missions and Warning indications A Any loud and or unusual noise from any part of the equipment should be taken as a sign of a problem and the equipment closed down and inspected immediately A Any smoke or smell of burning should be taken as a sign of a problem and the equipment closed down and inspected immediately A whistling noise could indicate a dry mechanical seal Stop the stirrer and lubricate the seal immediately Exchange if necessary In normal operation some additional noise and heat may be generated the extent depending on the phase of operation see performance data Service amp Maintenance gt Only fully qualified and authorised persons may repair the equipment Cleaning and routine maintenance information in provided in the main manual Before you begin Copyright You may not copy or duplicate any part of this manual without the prior written agreement of INFORS AG Switzerland Modification Due to the constant development and improvement of our products the bioreactor system supplied to you may differ from the one described in this manual INFORS explicitly retains the right to make such deviations and modifications Safety aspects A number of points relating to the safe operation of this machine and its proper use maintenance are contained in this manual It is vital you read all relevant sections before bringing the bioreactor into use and comply with the instructions given during its operat
74. n Click Menu entry F1 RUNNING lt lt Back Select 1 Stop Diagnose Selected fermenter 1 Parameter Feed Channel 6 Pump speed 0 Pump revolutions 0 Calibration start Pumped volume weight 0 Pumpfactor 424 Esc click Exit or value entry Enter F1 RUNNING lt lt Back Select 1 Stop Diagnose Selected fermenter 1 Parameter Feed Channel 6 Pump speed 50 Pump revolutions 0 Calibration start On RUNNING Pumped volume weight 0 51 F1 RUNNING lt lt Back Select 1 0 Stop Diagnose Selected fermenter 1 Parameter Feed Channel 6 Pump speed 50 Pump revolutions 30 Calibration start Pumped volume weight 0 Pumpfactor 1 Esc click Exit or value entry Enter 15 02 2008 Operating Manual Labfors 3 e page 58 of 79 F1 STOP 14 3 4 Diagnose Advanced Level 2 or 3 This menu indicates the status of pumps valves relays and other Selected fermenter 1 inputs outputs It is also possible to force the operation of valves Please take if using this menu as unauthorised interference can make the Auto AIR Auto Water bioreactor un usable Auto 02 Auto Over temp OK Auto c02 Auto Auto STIR Auto Type of stirrer is now also displayed here Bsti bacterial or CSti for cell Auto ALAR Auto culture Auto DO A Auto Auto DO B Auto The additional inputs and outputs allowed by the system are als
75. n FreeC 6 Analog 7 Anderungen Ersetzt durch Ersatz fir Extern I O steckerbelegung External Pin configuration Verz Datei IDatei Elektronik Corel Labfors3 DB25 Female maybe used internal All rights in particular the copyright in this document belong to INFORS AG Bottmingen SwitzerLand The document is given to you solely for your personal use according to the agreements Infors concluded with you every use which exceeds the above mentioned agreements including the transmission to third parties x90 70435 0704 cdr the making of copies Fotocopies extracts as well as the storage on data carries of any nature requires the prior written consent by INFORS AG IN FORS VW cousine 50704 VER
76. n depends on the configuration of the system At its simplest it is connection of the inoculation needle to the top pate by piercing membrane and overpressuring the vessel containing the inoculum to transfer it into the vessel The volume of inoculum is typically 5 10 of the working volume of the bioreactor 1 After autoclaving when ready remove port closure 2 Add ca 1ml of 70 ethanol to surface of membrane Use a Bunsen flame near to the port to create warm updraft OR set fire to the ethanol held in the collar 4 Pierce membrane with needle and screw into place Use gravity or overpressure to add inoculum 1 5 Clamp off line to inoculum vessel Optionally the inoculum can be concentrated into a small volume and added by syringe OR can be poured into the open port no membrane under a laminar flow cabinet INFORS 13 9 Aseptic Sampling Sampling takes place at various times after inoculation Usually a sample is taken immediately after inoculation to determine the initial number of organisms in the culture To use the sampling device a syringe and portable Bunsen burner are needed Spare bottles which have been previously sterilized are also required The size of sample should be determined by your analytical needs but also taking account of the volume of culture and the duration of the fermentation eg 100ml samples every 4 hours from an experiment using a 1 2L working volume vessel which is destined t
77. n the line Take to avoid injury due to needle sticks when handling the inoculation sampling and reagent delivery systems e Always work to GMP and or other appropriate safety standards e Only allow suitably trained operatives to use the bioreactor Please ensure a risk assessment is carried out if necessary INFORS NT 11 3 The vessel The vessels for Labfors3 bioreactor are normally delivered complete with internal fittings already in place drive shaft baffles impellor sample pipe and port closures Other items for fitting to the top plate will be packed separately eg exit gas cooler pH pO2 antifoam electrodes and inoculation needle Vessels come in several sizes covering the range of 2 13L total volume All are held in a metal support frame to which the reagent bottle holder can be fitted e The body of the vessel is borosilicate glass and the top plate plus fittings are constructed from 316L stainless steel All vessels have a round bottom e hey be equipped for bacterial or cell culture applications e he number and size of ports in the top plate varies with the size and type of vessel e Vessels can be jacketed to allow water circulation for temperature control or single walled for use with a heater pad and cold finger Connection to the water supply is via rapid coupling connectors if these are stiff on first use a little silicone grease applied to the male en
78. neral A NEVER open open or remove covers internal or external with the power switched on A No operations beyond those expressly stated in this guide are authorised by INFORS as being suitable for the equipment A All work on the equipment including adjustments repairs pipe couplings etc must be undertaken by professionally qualified staff A When repair and maintenance work has been completed any safety equipment provided must be refitted in its original state before the equipment 15 started A The equipment must be installed in accordance with the instructions contained in this user manual A The equipment s weight is over the permitted allowance of kilos pounds that people may lift so it must be lifted mechanically specifications sheet A Users are responsible for ensuring that the equipment 1 used in accordance with safety procedures applicable to their work and Is free of any biological or chemical contamination if an examination by INFORS staff is requested INFORS will not be held responsible for any equipment which has been improperly used maintained modified or repaired nor for any consequential losses arising A All the housing covers of the basic unit and operating panel are as they may cover critical areas only to be removed by personnel explicitly authorised by INFORS to do so If in doubt about any aspect of the use of this equipment or its suitability for an application please contact
79. o configured from this screen see Appendix Tun Ehe rotary 178 Sezoll Continue the rotary L R Scroll Continue To test an actuator function eg Acid pump e Click rotary switch to enter main part of the screen When the cursor is against Acid the key can be used to toggle the status from OFF ON AUTO e When Off the should not operate even if the pH controller requires acid e When set to ON the pump should run continuously e When set to auto the pump will only work when the pH controller requires acid If a pump is active The motor is shown as ON automatically Additional Indicators e Man Pump The push button on top of the pumps is shown as active on the display when pressed e Water OFF Water cannot be detected by the sensor in the thermo circulator and heating is therefore turned off running dry or distilled water has been used Overtemp OFF the 80 C over temperature cut out has been activated resets itself on cooling NOTE An additional mechanical over temperature device is fitted se to 105 C and this may have to be re set manually Check by opening the bottom panel of the base unit and pressing the red button on the thermo circulator 14 4 Parameter Submenu 14 4 4 Setpoint There are two ways to access this series of tabs 1 When the bioreactor is inactive via the Param tab and selecting a parameter name from the list Click to enter the main part of the screen Move the cursor bl
80. o last for 3 days is clearly not possible Typical sample volumes are in the area of 10 20ml for a typical bench scale bioreactor If the bioreactor is used in continuous mode it is possible to make a sample bypass in the harvest line and so allow the sample device port to be used for other purposes The sample device can also be used for inoculation if a longer dip pipe is inserted and the syringe used to blow the inoculum over 1 Remove any aluminium foil covering the air filter and connect a syringe eg 25 on the open end of the small filter 2 Loosen the clamp on the line and draw back on the syringe until liquid flows into the bottle 3 When 1 2 2 3 full push the syringe plunger down to blow the contents Attach of the sample line back into the vessel 4 Re clamp the line remove the bottle and cap it aseptically 5 Fita new bottle to the cap SCARE Sharp needles and screw into place 6 Forthe next sample draw a little culture into the bottle the DISCARD it Use a new sterile bottle to actually take the sample Pull Operating Manual Labfors 3 e AIT Arie NI a IM v JI m mo L 15 02 2008 13 10 Ending a fermentation Take care to follow this sequence when closing down for safety reasons e Ensure that services lines eg Gases cooling water etc are not pressurised Make sure that no medium can get into the gas supply system e Then initiate Closedown
81. ock to the required parameter and click 2 From the Values tab when the bioreactor is running Click to enter the main part of the screen Move the cursor block to the required parameter and click Use the Back tab to return to the main offline menu or Values tab The Setpoint tab is automatically highlighted The format is similar for most screens but can vary in the details according to the functions needed for individual parameters Explanations 1 Setpoint value in the parameter units 2 Alarm limits in whole parameter units This means eg 2 will cause an alarm for a pH 5 0 and 9 0 3 Regulation On shows the parameter is controlled by a calculation from the processor within the X DDC system 4 Output Active ON shows that control of the actuator pumps or valves is active Stirrer speed and Flow MUST have regulation switched OFF to function correctly as they send set points to controllers specific to those parameters Everything below Output Active is a special option for pO2 With the parameter pH Temperature Compensation is included This MUST be ON for pH to work properly during a fermentation With the parameter AF 1 Instead of Setpoint Dose Time Pump run time 2 Instead of alarm Lim Wait Time Pause between dose 3 An addition Regulation option ON OFF is provided If OFF it provides for IRIS control or manual with 0 100 this value being the percentage of dose tim
82. onal to the installation of the Acid Alkali and Antifoam pumps which operate automatically They are all digitally controlled This means that flow rates must be set through shot delay cycles of pump operation Set up of the Digital Feed Pump For the Feed pump the cycles can be set as 0 100 On time in seconds of the total cycle time with intervals of 0 1 seconds The pump can deliver up to 8 89ml min using the standard size of tubing provided with Marprene mid section of 2 79mmlD options available The pump can be calibrated from the Pumps tab The Dose time as a percentage 0 100 is set in the set point screen of the Feed parameter Ensure Regulation if present and Output Active are set to ON If a constantly variable flow rate is required it is better to work with a pump under analogue control Either the optional analogue pump must be fitted or an external analogue pump must be used with the analogue connector on the base unit 13 7 Other Possibilities There are other possible options displays and settings not covered in this part of the guide The information provided is sufficient for you to prepare sterilize and then operate the bioreactor with the minimum of delay Particularly in the case of fermentations of long duration it is usual to check the internal pH measurement for drift by comparison with an external measurement This can be corrected for using the Offset Value in the Calibration screen
83. our laboratory c Three gas and three plus one gas mix Connections are made to the base unit with the supplied cables and the gas bottles connected for oxygen nitrogen and carbon dioxide if fitted After configuration in the X DDC instrumentation to cascade gasmix to the dissolved oxygen controller the gas mix unit creates an artificial air mixture which is passed to the vessel at a constant flow rate via the mass flow controller A manual setting can be given eg 2096 oxygen or the process is automatically controlled based on the set point for dissolved oxygen The supply of carbon dioxide for pH control in place of acid is made separately to the main gas flow Carbon dioxide is added to the head space rather than entry via the sparger or special gassing system as standard Head space gassing with air oxygen is also possible as is the entry of CO via the sparger These and other possible options are on request 15 2 1 Detailed Description of the Infors Gas Mix System The gas mix from Infors can be configured according to requirements The standard configuration 1 suitable for all types of cell especially mammalian An air oxygen nitrogen mix is added 1 delivered directly to the air inlet filter of the sinter sparger in the vessel via a thermal mass flow control valve mounted on the side of the box Carbon dioxide is delivered to a separate inlet pipe in the head space above the culture liquid as a pulsed flow ac
84. overs a detailed description of the X DDC instrumentation Offline Menus amp Options Online Menus amp Options Parameter Options Description of control loops e The Appendices include as appropriate to your system Additional training notes experimental protocols amp practical training courses are available for users new to fermentation please contact us TABLE OF CONTENTS 1 CEconiormity CS CVA dee ae se sca 3 2 Quality Management 6 lt lt lt lt 3 E g 8 CI ON 4 5 1 Vessel identification Umm 4 E 5 6 1 C e 5 6 2 Safety points relating to installation amp 5 6 6 3 Safety points related to testing the CQUIDMENL ccccccceeeeeeeeceeccaeeeeeeceeeeesaeeseeeeessesessceeeesssueasaeasseeeeeesseaageess 8 7 Emissions Warning INGICATIONS P OE omm 8 COIN DETAILG ce esac tes 10 SEE LF ee ene 14 9 1 Delivery MOma UO
85. packed system over long distances a low trolley or palette raiser is recommended If the bioreactor is to be transported by a fork lift truck it is vital it is secured in such a way that there is no chance of it falling off or otherwise being damaged Packing materials The bioreactor and auxiliary equipment is shipped in a wooden crate All packing materials are environmentally friendly and can be recycled Unpacking and Checking Please make sure that no part of the bioreactor sustains damage during un packing Use the delivery documentation to ensure that all parts are present If there is a discrepancy between the list and the contents and or there are any signs of transport damage please contact your local Infors representative at once If this advice is not followed promptly any resulting costs will be the responsibility of the purchaser i e you 9 2 Location The bioreactor is best mounted on a laboratory bench of suitable working height For the size of space required see dimension drawings in Appendix Access to air water and power services should be nearby along with a suitable drain sink A free space of 50cm should be left all around the bioreactor to make servicing both easier and quicker The unit should be sited away from potential sources of electrical noise 9 3 Services The final connection of the bioreactor to the services will be made by INFORS if installation is requested by the customer This is aided gre
86. point of eg 200 rpm Click press to go from Navigation to For temperature ensure Measurement and Control Active fields are all switched ONFor antifoam Regulation ON OFF will be seen If ON then it main screen option works with the Foam probe if OFF then id can accept manual software dose times based on a percentage time of 0 100 active Enter numerical value 15 Click knob or press ENTER key to automatically start the fermentation and the Values screen shows actual values Switch a function ON or OFF or AUTO 16 You should hear see water rushing into the cold finger or vessel glass When full water will overflow and out to drain Confirm change and leave 17 The stirrer should now be active Allows return the tabbed area 18 Select Temperature using the cursor then Setpoint Enter a new value of the screen or aborts current of eg 30 C Carefully feel the jacket for signs of heating action 19 End the fermentation using the Stop tab and pressing 1 follow the instructions on screen Select bioreactor Choose parameter Fl STOP F2 STOP F1 STOP F2 STOP In 2 4 Config Passwrd Receipe lt lt Back Start Fe Pump Diagnose gt gt Selected fermenter 1 Fermi Temp 24 0 24 0 Stirrer Stirrer 180 7 0 2 7 4 0 00 Flow 2 70 0 4 Off Gasmix AFoam 0 00 gt 0 00 FreeA Flow 0 00 s s 0 00 FreeD Feed Off 0 00 FreeF Gas mix Off 0
87. r parameters Accept amp start fermentation When complete for all parameters pressing ENTER or clicking the rotary knob will automatically start the fermentation and the tab options change The bioreactor details for online operation will be shown on the Values screen A number of shortcuts are available Scroll cursor When a parameter has been selected using the rotary knob setpoints can snes be entered directly using the numeric keys E pH The key can be used to turn individual parameters On or Off Click a parameter for the full range of options Click to access parameter sub menu options i e Setpoint Calibration PID Pdiag F1 RUNNING 14 3 Values Screen 7 Online Operation lt lt Back Select 48 Pump Stop Diagnose Selected fermenter 1 Clicking to enter this screen will cause a block cursor to appear to the left of the fist parameter Clicking again will take you directly to the parameter submenu for that parameter Pressing Esc will take you back to the tabs Parameters not shown on the initial display can be reached by simply turning the rotary knob to the right until the screen scrolls upwards to reveal the remaining 8 parameters in turn Current set points are shown here along with the output from the controller If control is not active Off will be shown and the set point will be replaced with a series of dashes F1 STOP lt lt Back Start 24 84
88. r in option 4 x Peristalltic pump ubing used Marprene 64 shore D 2 79 x 4 39 biggest possible tube Speed Flow rates Temperature RPM as options pH pO2 AF gas flow level pressure weight feed analog pumps Max 16 parameters Vessel Dimensions i 1 0 115 240 1 D 115mm H 370mm 1 0 150mm H 235mm 1 0 150 H 465mm 1 0 200 352 200 420 Glass borosilicate Cylindrical with round bottom Removable support with 3 baffles 12 19 round thread 2x4mm fix weldet 10mm one Pt 100 DC 0 5Nm 0 5 Nm DC 0 5 Nm DC 0 5 Nm 2 5 Nm 0 5 Nm Servomotor 1 4Nm Power 150 W 150 150 W 150 W 100 150 W 680W Range with 2 Imp 80 1500 Cell 20 300 80 1500 20 300 80 1500 Cell 20 300 80 1400 Cell 20 300 Cell 20 300 80 700 10 1250 Cell 20 300 earing Bacterial Bearing housing with mechanical seal earing Cell Guide and bearing in Vessel x Impeller Optional 3 type Rushton with six blade Impellers pe Outer Diameter D 46 12 D 46mmxi2mm D246mmx12mm D254mmx12mm 70 12 D 70mm x 12 mm Temperature T 100 10 to 150 C ccurac 0 2 in range 10 to 60 C nlet Filter Sterilizable absolute 0 2 um filter Outlet Filter Sterilizable absolute 0 2 or 0 45 um filter Control x Rotameter additional Rotameter Mass flow controller in option ccurac Rota
89. rilization of a Cell Culture system 13 1 2 After autoclaving the vessel must be allowed to cool to a comfortable handling temperature before replacing it on the base unit of the bioreactor Cooling down and replacing the vessel It will be several hours before the electrolyte within the electrode cools despite what the overall vessel temperature may fall to This time can be used for polarizing the pO electrode by connecting it to its cable grey lead and leaving the instrumentation switched on for at least 6 hours The temperature control for the vessel can be switched on AFTER the following procedures have been completed 1 The vessel is located on the bioreactor base 2 heater pad is wrapped around the body of the vessel and secured OR 3 The rapid coupling connectors on the vessel jacket are connected to the base unit pipes Tubing is connected to the overflow pipe and run into a drain 4 The pipes for the exit gas cooler and or cold finger are connected to the water supply and drain 5 The main water tap or circulator is switched on 6 The inlet valves for water exit gas cooler supply are turned ON and adjusted for flow rate The stirrer is placed on the vessel drive hub and turned until it locks into position Final Checks before Autoclaving VESSEL 2 Exit gas line has a 0 45u filter fitted and is OPEN Inlet gas line sparger has a 0 2u filter fitted All unused ports are fitted with a closure
90. rite protect tab on the back edge of the card is set towards the middle of the card in the WRITE position not protected before replacing it 5 The card will be automatically initialised on starting the bioreactor This will take a few seconds 2NB A memory card MUST be present for the bioreactor to operate Never start the unit without a card in place Memory card battery holder battery 9 7 Maintenance points We recommend that all O Rings and Membranes should be changed at least once a year Clearly any deformed or damaged O rings must be replaced before each run Before fitting the O ring wet the O ring with alcohol Pierced membranes must be replaced During operation any spillages of acid or alkali should be cleaned up immediately At the end of the fermentation and emptying of the vessel and reagent tubes the vessel should be sterilized and rinsed several times with water 0 1 N NaOH is recommended for denaturizing proteins 2 Observe all safety precautions regarding skin or eye contact and inhalation of such chemicals as defined by your local regulations regarding the treatment of hazardous chemicals 9 8 Mechanical seal The mechanical seal must be replaced at regular intervals The seal is destroyed by running dry Therefore it is vital to ensure this component is always supplied with fluid glycerine See instructions on the next page for topping up the seal 9 8 1 Electrodes The pH and pOz
91. s and fitting are placed in the holder which is hooked onto the pins of the vessel support frame and the whole arrangement sterilized together with the vessel in an autoclave Operation use and cleaning of corrective reagent tubing and pumps gt WARNING do not autoclave the reagent bottle lines filled with corrective agent as this could damage the dip tube Use water instead Take care regarding overpressure Tubing can burst or come off the inlet pipes 2 As part of the cleaning regime the corrective reagent tubing MUST be rinsed with water Storage and autoclaving of tubing filled with acid or base can result in damage to the tubing and also represents a safety hazard to the user INFORS NT 11 7 Sample Device This is prepared in the same way as the reagent bottles except a short pipe with a needle end replaces the dip tube A number of spare capped empty bottles are prepared and autoclaved ready for use when sampling How many bottles you need depends on the number of samples you wish to take but always prepare double what is required to allow for discarding pre samples of culture The metal holder is screwed in place on the top plate support and a silicone tube connects the inlet to the sample bottle and the dip tube already located in the vessel The line is clamped off for autoclaving and remains closed until a sample is to be taken The use of the sampling system is covered later in this manual 11 8 Pr
92. s flasks are used The top plate of the flask is always fitted with a dip tube and some form of pressure relief system Either a metal top plate or a membrane may be in place Choice of reagents and their strength varies according to which microbes are being cultured Refer to literature 2 Do not use hydrochloric acid and ammonia must not be autoclaved explosion risk Fill the 250ml reagent bottles approximately 2 3 full with either acid base or antifoam as appropriate Fit the bottle cap using a silicone membrane to seal the open top Prior to sterilization fit a pressure relief outlet using the disposable needle and small disposable filter from the Service Set The pump tubing sized to be long enough to reach from the final location of the container via the pump head and to the vessel inlet on the top plate Use a Marprene insert in the area compressed by the peristaltic pump rollers for maximum tubing life An example line is prepared in included in the Service Set Connects the tubing to the dip tube long needle in the reagent bottle The outlet from the flask dip tube is closed off during autoclaving due to pressure differences between the inside and outside of the flask With glass flasks the pump tubing must be clamped off with a suitable clamp eg a gate clip The reagent bottles are now set up already connected to the inlets of the 2 way or single needle fittings and simply clamped off for autoclaving The bottle
93. s not fulfilled 9 9 2 Disposal The LABFORS bioreactor is mainly constructed from metal with some glass plastics and electronic components Disposal by normal methods is possible along with the motor heaters and other internal components Please ensure that all necessary steps have been taken to ensure microbiological safety before considering disposal 9 9 3 Identification plate and serial number The ID plate and serial number can be found on the right side of the base unit INFORS 10 A Test Run Before preparing the system for first use a functional check on the services and basic operation of the stirring temperature and gas supply systems can easily be carried out Full details of the individual procedures can be found in the later sections 1 Ensure that the mains switch on the base unit is OFF and the manual water inlet valves at the right bottom of the base unit Green knobs are turned off fully clockwise Likewise the rotameter needle valve should be fully closed black knob below the glass rotameter scale Air Inlet at right rear 2 Check all connections to services lines are tight and that air and water are turned on at the supply CHECK FOR LEAKS 3 Fill the vessel to the working volume with water and then connect to the base unit Exit Gas Cooler Water Inlet Exit Gas Cooler Valve Water Supply IN Water inlet valve to Jacket Double jacket Water outlet from double jacket to base unit
94. t 15 02 2008 Operating Manual Labfors 3 e page 75 of 79 NW150 2 3 3 6L 2 CL MELE ESL 1L min WV VU EEE x kk P 77222272 72 7727774772777777777777777177772 155 2 2L min WV page 76 of 79 Q gt c Operating NW150 5 7 5L 2 2L min WV AAT Ll d nd ne iP i i n i iP iP i iP i p aa LO drca a ai 0 2 7L WV zd UFFEERFEREREFREREFEFREFEFRFREFAFREFEFEREREEEFREREFEERIEEFREPEIKEERIFAFAERIKEFRERER Fu PA cm i MC INFORS 15 7 Specifications INFORS HT Labfors 3 Fermentor Labfors 3 Specifications _ 3sttes aetiters 7stiters toners nues Fermentor Dimensions W D H 475 x 475 x 880 mm 475 x 475 x 880 mm 475 x 475 x 880 mm 475 x 475 x 880 mm 475 x 475 x 880 mm 475 x 475 x 880 mm Weight Base Unit Electrical 230V 10A single phase supply Integrated In Outputs 4 x Analogue input 0 5V 0 10V 0 20 4 20mA 6 x Analogue output 0 20mA 4 20 2 x digital output open collector Electric heating up to 95 C thermo cirqulation 70 C max and tap water cooling heat colling programmable 5 utility water to 70 C Chille
95. t An information screen Ferm shows eem The operating panel is connected to the base unit via a cable with twin RJ45 connectors running from the back of the unit to a socket at the rear of the base unit Ensure this cable is securely plugged in before starting the unit The screen backlighting automatically switches off approx 30 minutes after the last key press The display is then dark The backlight activates again as soon as any key is pressed If desired the backlight can be switched on permanently using Menu Options Rotate to move position of cursor highlight tabs Press to go from Navigation to main screen option Enter numerical value Switch the function ON or OFF or AUTO Confirm change and leave menu option Acid pump On Base pump On Feed pump On Foam pump On Heater On Cooling On Water pump On Coms status OK Water present Unused Mains Power ON Leaves menu option without making any changes Enter Allows return to the tabbed area of the screen some exceptions eg Diagnostics Please note the following basic guidelines 1 Numeric values eg Setpoint pH 7 2 must be confirmed by Enter in Calibration mode With a second Enter the change is accepted and you leave the menu option For other numeric fields moving the cursor along is sufficient 2 Switching On and OFF eg Output active ON OFF does not require confirmation by Enter Pressing the Enter
96. ter i Fl Z2TOP F2 5TOP 14 2 Selected Bioreactor fermenter o LO For a single bioreactor click to enter the main screen For multiple bioreactors first select the unit number required 26 0 When an individual bioreactor is selected additional options become d 0 00 available for offline set up namely 80 0 0 00 0 00 Off 14 2 1 Start Off For multi bioreactor operation up to 4 Labfors3 bioreactors can be controlled from a single X DDC operation panel Consequently the number of the bioreactor to be started first has to be selected The Start screen can Switching parameters on amp then be accessed starting the fermentation Click the rotary knob to move to the main part of the display Turning the knob will now let you move up down in each column of parameter names Start with the order being temp stirrer pH pO2 etc Use the key to toggle between Off and the current setpoint for each Y parameter equivalent to switching the parameter On Entry of set points 2 is done using the Param tab prior to starting the bioreactor or directly by entering numbers into the OFF field for a parameter Previously entered set points from a completed fermentation will be stored pH Off and appear at this point If your bioreactor has been under remote control it will be the last external set point given which will have been stored 4 pH 7 0 Repeat for othe
97. the parameter 9 Output ramp A slow stepwise activation of an actuator can be activated here e g Stirrer speed or mass flow control valve action 10 D Term Differential term D Accounts for the rate of change of the error over time and compensates for this to limit any overshoot In most cases it is not needed 11 Dead Band Inside the dead band range Setpoint given value the controller is inactive 12 Step size This value is given as a 96 of the selected controller out put The control output will then be increased by this amount at regular intervals the Value in Eval Time in seconds 13 Neg Factor If a controller has two actuators of differing strengths e g strong acid weak alkali this value can compensate by adding a weighting factor or bias to the weaker controller output 14 Eval Time This value gives the time interval in seconds between re calculation of the PID equation and hence changes in the controller output 10 seconds is a good average value INFORS PID P is actually the Gain P 100 gain the bigger the value the sharper the control l 1 sec hence a small number as increase decrease has a large effect amp D is set in seconds often zero You should only alter these settings if you are confident about using PID control and the precise sequence and consequences of any changes can be evaluated The PID control is governed by the following formula Error Set Act
98. urrent level is editable Explanatory note to level 7 It is not possible to deactivate the pincode for level 7 A fixed INFORS in house pin code exists additional to the Administrator pin code Alteration to level 7 is always possible as soon as a password pin code is asked for on the display Setting Change pin to ON the pin definition display shows the following information New Pin Pin change of level Current Pin code New pin code Repeat Pin code KKKK KKKK Only level 7 allows a Pin code change Change Pin Users with lower levels can neither change this nor anyone else s Pin code Basically every function that needs a pin code will automatically and visibly ask for it on the display This is not dependent on active or inactive sequences Pin protected domain Level 1 2 3 7 Pin lowest needed level is automatically shown after Level on the display General view access authorisation Standard user level 1 Start Set points Calibration only pH and pO2 Advanced user level 2 Diagnose Advanced user level 3 Recipes PID settings Configuration A detailled scheme can be provided on request 14 1 5 Recipes advanced level 3 The Recipe tab allows access to the various communications and configuration storage options available within X DDC Simply press the number of the required option to go directly to that screen The entire information re
99. usandoadaensenseaeeaidebusadardeaedeisancundgupsaianeaasaieiceues 27 111 Checking Services Connections ccccccceeccccccececeeeeeeeceeeeeeeeeeceeeeessaeeeeeeeeeseeeeeeeeeeessseeeseceeeseeeesesseaaeeeeeeeeesaas 28 UE MEC e mm doc 28 1 2027775 29 11 4 Switching ON at the base unit and 30 11 4 1 X Starting a fermentation E RaT rns snas 31 E 32 11 5 1 Preparation and calibration of pH 32 11 5 2 Dissolved Oxygen pO2 Electrode preparation 34 11 6 Reagent bottle 34 DOY E DONNE EC EE 35 11 5 E Preparations 35 00 1900 etm mm ee 35 11 8 2 Inoculation 35 L83 36 iE E c E A 36
100. w growth rate and undemanding oxygen requirements Gas flow can be set to a constant value manually or changed according to the dissolved oxygen level In this latter case setting a maximum on the rotameter as well as passing the gas through the mass flow controller will ensure that the system is not gassed at too high a flow rate Setting of the gas flow controller is covered in the next section b Oxygen only or Two gas supplementation Bottled oxygen and carbon dioxide gases are connected directly to the relevant valve with flexible pressure tubing with suitable flow meters and pressure reducers connected to the gas bottles An air pressure of 0 5 0 9 bar and a flow of eg 0 1 vessel volumes per minute VVM ensures the addition of each gas does not cause problems when it blends with the main air flow from the rotameter even if both valves work simultaneously Set the main air flow via the rotameter typically for 0 1 to 0 3 VVM maximum and or adjust the mass flow controller to this value used Once configured control of the valves is automatic from the dissolved oxygen and pH controllers For pH the acid pump will operate at the same time as the gas valve This allows maximum flexibility of use if a non gassed pH control is needed for different tasks in the future 2 Inlet pressure should be reduced to 2 0 bar using a pressure reducting valve Be sure that you follow any standing instructions regarding the use of O2 and CO2 in y
101. y already be fitted Outlet pipe to sparger Glass tube with scale L min bottom of ball Needle valve adjust Sparger inlet 2When using a Mass Flow Control Valve for gas flow option If the Mass Flow Control Valve is activated without first turning on the gas supply a huge control error will build up This influences any adjustments necessary by the valve to reach the pre selected set point for flow rate When the gas supply is later turned on this error causes the set point to be exceeded by a large margin which is clearly undesirable gt Therefore always turn the gas supply on before setting Output Active to ON in X DDC or otherwise activating the Flow parameter 8 Open the Water Inlet Valve BOTTOM green knob so water can flow 9 Insert Pt 100 temperature sensor into the pocket in the top plate until metal to metal contact can be heard and or felt orange cable with metal sensor end permanently attached A Preview of the Operating Panel 10 The Operating panel will have completed its self test and you will see the FERM screen 11 Select bioreactor 1 4 if applicable 12 Use the rotary knob to go to the START tab click to select then turn the knob to move the cursor to Temperature and click the knob T Rotate to move position of cursor 13 Type in a set point of 5 and press 2 x the ENTER key To navigate in menu Highlight tabs 14 Repeat this process for Stirrer and give a set

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