Infors HT Minifors 2 Operating Manual
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Infors HT Minifors 2 Operating Manual

Benchtop bioreactor
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Operating Manual
2019-02-07
Minifors 2
Benchtop Bioreactor
Dok-ID: 75623 / V.03.00 - Original

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Summary of Contents for Infors HT Minifors 2

  • Page 1 Operating Manual 2019-02-07 Minifors 2 Benchtop Bioreactor Dok-ID: 75623 / V.03.00 - Original...
  • Page 2 Infors GmbH Infors UK Ltd Infors Dachauer Str. 6 The Courtyard Business Centre Headoffice, Schweiz D-85254 Einsbach Dovers Farm, Lonesome Lane, Rittergasse 27 T +49 (0)8135 8333 Reigate CH-4103 Bottmingen F +49 (0)8135 8320 Surrey, RH2 7QT, UK T +41 (0)61 425 77 00 infors.de@infors-ht.com T +44 (0)1737 22 31 00 F +41 (0)61 425 77 01...

  • Page 3: Table Of Contents

    Minifors 2 - Benchtop Bioreactor Table of Contents General Information ............... 9 About this Manual ............9 Explanation of Special Notices ........9 1.2.1 Warning Notices ..........9 1.2.2 Other Notices..........10 Equipment Identification (Standard Identification Plate) ..................10 Declaration of Conformity ..........11 Customer Service and Services ........
  • Page 4 Minifors 2 - Benchtop Bioreactor Table of Contents 3.1.10 Connections for Sensors (Sensor Cables) ...25 3.1.11 Sparger Connection (Gassing) .....25 3.1.12 Connections and Water Flow Control Valve for the Exit Gas Cooler .........26 Operating Panel ............27 Culture Vessel ..............27 3.3.1 Top Plate ............29...
  • Page 5 Minifors 2 - Benchtop Bioreactor Table of Contents Electrode Holder ............49 Addition Port Adapters and Feed Needles ....50 Septum Collar .............. 51 Dip Tubes ..............51 5.10 Pocket for Temperature Sensor (Pt100) ...... 52 5.11 Exit Gas Cooler ............53 5.12 Reagent Bottles ............
  • Page 6 Minifors 2 - Benchtop Bioreactor Table of Contents 8.1.3 Mounting Dip Tubes and Spargers ....76 8.1.4 Inserting the Vessel into the Vessel Holder ..77 8.1.5 Inserting the Baffles ........78 8.1.6 Moistening/Filling the Culture Vessel ...79 8.1.7 Fitting the Vessel Top Plate ......79 8.1.8...
  • Page 7 Minifors 2 - Benchtop Bioreactor Table of Contents 8.2.10 Connecting the pO Sensor ......106 8.2.11 Calibrating the pO Sensor ......107 8.2.12 Checking the Hoses and Hose Connections ..............107 Cultivation ................108 Preparing the Medium ..........108 Sampling ..............109 Inoculation ..............
  • Page 8 Minifors 2 - Benchtop Bioreactor Table of Contents 10.4.4 ..............143 10.4.5 TotalFlow ............146 10.4.6 GasMix ............146 10.4.7 Foam ............147 10.5 EXTENDED Parameter Group ........149 10.5.1 Turbidity (Optional) ........149 10.5.2 Balance (Optional) ........149 10.5.3 AirFlow ............149 10.5.4 Gas2Flow ............150 10.6 EXITGAS Parameter Group ........151 10.6.1...
  • Page 9 Minifors 2 - Benchtop Bioreactor Table of Contents 11.3.1 Removing the Exit Gas Cooler ....185 11.3.2 Removing the Sensors ....... 185 11.3.3 Removing Hoses, Filters and Pump Heads ..............186 11.3.4 Removing Blanking Plugs ......187 11.3.5 Removing the Septum Collar and Septum . 187 11.3.6...
  • Page 10 Minifors 2 - Benchtop Bioreactor Table of Contents 14.4.3 Water OUT, Basic Unit .......207 14.4.4 Gas (Air, O or N ) ........208 14.5 Specifications .............208 14.5.1 Operating Panel ..........208 14.5.2 Culture Vessels...........208 14.5.3 Stirrer ............209 14.5.4 Temperature ..........210 14.5.5 Gassing ............211 14.5.6...

  • Page 11: General Information

    Minifors 2 - Benchtop Bioreactor General Information 1 General Information 1.1 About this Manual This manual enables the safe and efficient handling of the equip- ment. All the information and instructions in this operating manual comply with the current standards, legal regulations, the latest technologi- cal and scientific developments and the knowledge gained from the manufacturer’s many years of experience in this field.

  • Page 12: Other Notices

    Minifors 2 - Benchtop Bioreactor General Information WARNING The signal word “WARNING” indicates a potentially dangerous situation that may result in severe or even fatal injuries if not avoided. CAUTION The signal word “CAUTION” indicates a potentially dangerous situation that may result in minor injuries if not avoided.

  • Page 13: Declaration Of Conformity

    Minifors 2 - Benchtop Bioreactor General Information 1.4 Declaration of Conformity The equipment is in compliance with the essential requirements of the following Directives: Machinery Directive 2006/42/EC  EMC Directive 2014/30/EU  The Declaration of Conformity according to EC Machinery Directive 2006/42/EC, annex II 1 A is included in the general documentation supplied with the equipment.

  • Page 14: Safety And Responsibility

    2.1 Intended Use, Incorrect Use and Misuse The bench-top bioreactor Minifors 2 from INFORS HT is de- signed especially for research and development and for the cultivation of microorganisms in a biotechnology laboratory.

  • Page 15: Qualified Personnel

    Minifors 2 - Benchtop Bioreactor Safety and Responsibility For use for special applications not covered by conventional, in- tended use, the equipment must be modified and certified accord- ingly by the manufacturer. Any use of the equipment outside of a biotechnology laboratory, i.e.

  • Page 16: Operator

    Minifors 2 - Benchtop Bioreactor Safety and Responsibility Qualified personnel On account of their specific education, training and – in many cases – experience, the qualified personnel required for this work are able to recognise risks and respond accordingly to potential hazards.

  • Page 17: Unauthorised Persons

    Minifors 2 - Benchtop Bioreactor Safety and Responsibility Technicians in training Persons in this group who are undergoing training or apprentice- ships are only permitted to use the equipment under supervision and in accordance with the instructions of a trained and qualified technician.

  • Page 18: Electrical Current

    Minifors 2 - Benchtop Bioreactor Safety and Responsibility The following notices are general in nature. As such, with a few exceptions they are not repeated in the remaining sections. 2.5.1 Electrical Current The equipment runs on electrical power. There is an immediate risk of fatal injury if contact is made with live parts.

  • Page 19: Hot Surfaces

    Minifors 2 - Benchtop Bioreactor Safety and Responsibility 2.6.1 Hot Surfaces For processes that are carried out with temperatures over 55 °C, there is a danger of burns on hot surfaces. Since the equipment is intended for applications at high tempera- tures, it is the responsibility of the users to ensure that they have sufficient protection.

  • Page 20: Bioactive Substances Or Pathogenic Organisms

    Minifors 2 - Benchtop Bioreactor Safety and Responsibility Since the equipment is used deliberately for such applications, it is the responsibility of the users to ensure that they have sufficient protection. 2.6.5 Bioactive Substances or Pathogenic Organisms The use or production of bioactive substances, pathogenic organ- isms or genetically modified cultures entails a significant health risk.

  • Page 21: Declaration Of Decontamination

    Minifors 2 - Benchtop Bioreactor Safety and Responsibility 2.8 Declaration of Decontamination When returning the equipment for repair, disassembly or disposal, it is required for the safety of all parties involved and because of legal provisions that a lawful declaration of decontamination is pre- sent.

  • Page 22: Setup And Function

    Minifors 2 - Benchtop Bioreactor Setup and Function 3 Setup and Function 3.1 Basic Unit Pumps LED signal strip Thermal block and adapter Main switch Connections for sparger and exit gas cooler Connections for sensors Operating panel Hooks for vessel holder All of the measurement and control technology is built into the basic unit.

  • Page 23: Main Switch

    Minifors 2 - Benchtop Bioreactor Setup and Function block plus adapter for regulating the temperature of the culture vessel, four pumps for adding reagents and feed solution, and the operating panel. 3.1.1 Main Switch The main switch is located on the right-hand side on the basic unit.

  • Page 24: Identification Plate

    Minifors 2 - Benchtop Bioreactor Setup and Function The pump drive shafts are located on the left-hand side of the basic unit. The drive shafts’ direction of rotation is set as standard to anti-clockwise for “filling”; see marking on mounting plate. The pumps can be configured individually using the operating panel, and thus each set to digital or analogue operation as required.

  • Page 25: Power Connection

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.1.5 Power Connection The connection socket for the power cable is located on the rear of the basic unit, at the bottom left. 3.1.6 Water Connections The water connections are located on the rear of the basic unit, at the bottom right.

  • Page 26: Signal Connections

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.1.8 Signal Connections The following signal connections with the corresponding symbols are located on the rear of the basic unit, at the bottom left: SERVICE: 9-pin RS232, for connecting a diagnostic com- ...

  • Page 27: Connections For Sensors (Sensor Cables)

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.1.10 Connections for Sensors (Sensor Cables) The basic unit is equipped as standard for measuring temperature, pH, pO and antifoam. This means that the temperature (Pt100) and the connecting cables for the pH, pO and antifoam sensors are always present.

  • Page 28: Connections And Water Flow Control Valve For The Exit Gas Cooler

    Minifors 2 - Benchtop Bioreactor Setup and Function The hose used to connect the sparger is connected to the basic unit’s gassing connection at the factory. 3.1.12 Connections and Water Flow Control Valve for the Exit Gas Cooler The water connections for the exit gas cooler are located on the front of the basic unit, at the bottom left.

  • Page 29: Operating Panel

    Minifors 2 - Benchtop Bioreactor Setup and Function The water flow control valve is located on the rear of the basic unit and marked with a corresponding symbol: The control valve is set at the factory. If necessary, the water flow rate can be set manually using the control valve.
  • Page 30 Minifors 2 - Benchtop Bioreactor Setup and Function Motor coupling Pump holder Vessel holder handle Reagent bottle holder Glass vessel Vessel holder Vessel holder stand Top plate The illustration shows a culture vessel with a total volume of 1.5 L and a nominal width of 90 mm.

  • Page 31: Top Plate

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.3.1 Top Plate Knurled screw (x4) O-ring Top plate Damping ring (spacer ring) Vessel Flange The top plate is attached to the vessel using four knurled screws and a flange. The knurled screws also hold the vessel in place in the vessel holder.

  • Page 32: Ports In The Vessel Top Plate And Their Configuration

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.3.2 Ports in the Vessel Top Plate and their Configuration The vessel top plate has different ports of different sizes to mount the different components such as sparger, blanking plugs, sensors etc. The number of ports in the top plate and its configuration de- pends on the diameter nominal (= inner diameter) of the culture vessel.

  • Page 33: Vessel Top Plate, Nominal Width 115

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.3.4 Vessel Top Plate, Nominal Width 115 Ø 12 mm Pg13.5: pH sensor Ø 12 mm Pg13.5: Additional sensor Ø 12 mm Pg13.5: Exit gas cooler Ø 10 mm: Temperature sensor (Pt100) Ø...

  • Page 34: Vessel Top Plate, Nominal Width 145

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.3.5 Vessel Top Plate, Nominal Width 145 Ø 12 mm Pg13.5: Exit gas cooler Ø 10 mm: Temperature sensor (Pt100) Ø 12 mm Pg13.5: Additional sensor Antifoam sensor earth connection Ø 7.5 mm: Addition port adaptors (4x) 10 Ø...

  • Page 35: Temperature Control System

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.4 Temperature Control System The temperature (heating and cooling) is controlled using a thermal block and its adapter. Thermal block adapter Thermal block Fastening screw (Allen screw, 4 pieces) Hook, 2 pieces There is a thermal block adapter for each vessel size.

  • Page 36: Stirrer

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.5 Stirrer The stirrer shaft is driven from above and turns anti-clockwise (left when viewing vessel from above). Drive hub Stirrer shaft Mechanical seal The stirrer shaft is sealed using a mechanical seal.

  • Page 37: Gassing System

    Minifors 2 - Benchtop Bioreactor Setup and Function A brushless gear motor with a mechanical coupling is used as standard. Depending on the size of the vessel, two motors with dif- ferent power levels are used; see the “Technical data” section, sub-section “Stirrer specifications”.

  • Page 38: Gas Entry

    Minifors 2 - Benchtop Bioreactor Setup and Function 3.6.1 Gas Entry A silicone hose leads the gas or gas mixture from the gassing con- nection on the basic unit to the culture vessel, via a sterile filter. The gas is fed directly into the medium via the sparger (sparger gassing).

  • Page 39: Po Control

    INFORMATION The matching pH sensor is included in the standard package. These pH sensors are pre-configured by the equipment manu- facturer, INFORS HT. Replacement sensors must be configured before use. For details on the technical data, maintenance and storage require- ments for the pH sensors, see the separate documentation pro- vided by the sensor manufacturer.

  • Page 40: Measurement System

     INFORMATION The HAMILTON pO sensors are pre-configured by the equip- ment manufacturer, INFORS HT. Replacement sensors must be configured before use. For details on the technical data, maintenance and storage require- ments for the pO sensors, see the separate documentation pro- vided by the sensor manufacturer.

  • Page 41: Antifoam Control

    Minifors 2 - Benchtop Bioreactor Setup and Function For more details on the electrode holder, see the “Accessories” section, sub-section “Electrode Holder”. 3.9 Antifoam Control Foam hinders the exchange of gas between the medium and the gas phase in the head space. The exit gas filter can become clogged with foam, which causes a pressure build-up in the vessel.

  • Page 42: Options

    Minifors 2 - Benchtop Bioreactor Options 4 Options The following options are available in addition to the equipment included in the scope of supply for the basic unit. 4.1 Turbidity Measurement The Optek ASD12-N measurement sensor can be used to deter- mine the turbidity of the culture.

  • Page 43: Sensors

    Minifors 2 - Benchtop Bioreactor Options 4.1.1 Sensors Optek ASD sensors are high-precision turbidity sensors. They can be used to draw conclusions regarding the growth of microbial cul- tures or cell cultures as a function of NIR absorption. Measurement gap (optical path length, OPL) Sensor head Shaft, Ø...

  • Page 44: Mounting The Sensor

    Minifors 2 - Benchtop Bioreactor Options This can be done on the operating panel, either before or after autoclaving, depending on the application in question. For more details, see the main chapter “Operation” chapter “Calibrating the Turbidity Sensor”. 4.1.3 Mounting the Sensor For culture vessels with nominal widths of 90 and 145, Optek ASD sensors can be mounted directly into 12 mm/Pg13.5 ports.

  • Page 45: Interferences Turbidity Measurement

    Minifors 2 - Benchtop Bioreactor Options 4.1.5 Interferences Turbidity Measurement Interference Displayed measured value is not plausible / unusual Possible Cause Remedy Sensor cable is twisted or kinked or Check and ensure that the sensor cable is not Operator not properly connected.

  • Page 46: Exit Gas Analysis

    Minifors 2 - Benchtop Bioreactor Options 4.2 Exit Gas Analysis In order to allow the user to draw conclusions regarding the status of the culture while the bioprocess is still underway, the CO measurements are often taken and analysed in the exit gas flow of the bioreactor.

  • Page 47: Calibrating The Gas Sensor

    Minifors 2 - Benchtop Bioreactor Options Connecting the sensor cable The fixed sensor cable is pre-installed in the factory (rear of equip- ment). The cable has an 8-pin round plug connector. In order to connect the sensor, the plug connector is plugged into the socket marked Port A on the gas sensor.

  • Page 48: Accessories

    Minifors 2 - Benchtop Bioreactor Accessories 5 Accessories The table below lists all the accessories included in the standard package, divided according to vessel size (TV = total volume) and nominal vessel width (inside diameter). Accessories 1.5 L TV/NW 90 3.0 L TV/NW 115...

  • Page 49: Sparger

    Minifors 2 - Benchtop Bioreactor Accessories It must be plugged into the opening of the drive hub for autoclaving of the culture vessel! 5.2 Sparger The gas is fed directly into the medium via a ring sparger (Ø 6 mm). The sparger is mounted in a 10 mm port in the vessel top plate using a clamping adaptor, and connected to the gassing sys- tem on the basic unit via a silicone hose with a sterile filter.

  • Page 50: Blanking Plugs

    Minifors 2 - Benchtop Bioreactor Accessories 5.4 Blanking Plugs Blanking plugs are used to seal open ports. There are different blanking plugs for the different types of port. Blanking plug, Ø 10 mm Fitted with fixed O-ring. A fastening screw is used to fasten it in the 10 mm vessel top plate port (see the “Clamping Adaptors and Fastening Screws”...

  • Page 51: Electrode Holder

    Minifors 2 - Benchtop Bioreactor Accessories Clamping adapter Ø 6/10 mm Fitted with two fixed O-rings (B & C). When the slotted-head screw is undone, the component with a di- ameter of 6 mm can be inserted in or removed from the clamping adapter.

  • Page 52: Addition Port Adapters And Feed Needles

    Minifors 2 - Benchtop Bioreactor Accessories 5.7 Addition Port Adapters and Feed Needles Addition port adapters and feed needles are used to feed liquid into the culture vessel. They each come with a tubing connection, are fitted with a fixed O-ring and are mounted into the four 7.5 mm ports in the vessel top plate.

  • Page 53: Septum Collar

    Minifors 2 - Benchtop Bioreactor Accessories 5.8 Septum Collar The septum collar with inside thread is used to inoculate the cul- ture in combination with the syringe, injection needle and septum (inoculation membranes); see the “Inoculation Accessories” sec- tion. The septum collar is used to hold the septum in place in the 12 mm/Pg13.5 port.

  • Page 54: Pocket For Temperature Sensor (Pt100)

    Minifors 2 - Benchtop Bioreactor Accessories 5.10 Pocket for Temperature Sensor (Pt100) The pocket is a pipe with a sealed bottom end, and is used to in- sert the temperature sensor. Pocket Ø 10 mm Fitted with fixed O-ring. A fastening screw is used to fasten it in the 10 mm vessel top plate port.

  • Page 55: Exit Gas Cooler

    Minifors 2 - Benchtop Bioreactor Accessories 5.11 Exit Gas Cooler The basic unit supplies cooling liquid to the exit gas cooler. The cooling liquid flow rate can be adjusted using the control valve on the basic unit. The two hoses for water supply (bottom) and return (top) are connected to the basic unit at the factory and simply con- nected to the exit gas cooler via the two rapid couplings.

  • Page 56: Reagent Bottles

    Minifors 2 - Benchtop Bioreactor Accessories 5.12 Reagent Bottles Two sizes of borosilicate reagent bottle are available for adding re- agent and feed solution: 250 mL  500 mL  Reagent bottles are fitted before delivery. There are two hose con- nections on the lid.
  • Page 57 Minifors 2 - Benchtop Bioreactor Accessories Content of the set The set consists of a completely pre-assembled group of valves with hoses and two syringes. It is connected via silicone hose with a dip tube. Valve assembly Sterile filter Check valve...
  • Page 58 Minifors 2 - Benchtop Bioreactor Accessories Principle of function The sample valve on the side arm of the T-piece opens by putting the Luer connector of the syringe into the valve and closes by re- moving the syringe. No further handling is necessary.

  • Page 59: Pump Heads

    Minifors 2 - Benchtop Bioreactor Accessories sample has to be processed under sterile conditions. For sampling, the same non-sterile syringe can be used repeatedly, without fear of contamination of the culture vessel. Aseptic Sampling For each sample, use a new, sterile syringe with Luer Lock fitting, in order to ensure the sterility of the sample.

  • Page 60: Vessel Holder With Built-In Holder For Reagent Bottles And Pumps

    Minifors 2 - Benchtop Bioreactor Accessories 5.15 Vessel Holder with Built-in Holder for Reagent Bottles and Pumps The vessel holder frame has a holding device for four reagent bot- tles and a holder for the four pump heads. Reagent bottle holder...
  • Page 61 Minifors 2 - Benchtop Bioreactor Accessories All the sterile filters in the scope of supply are autoclavable, dispos- able filters with PTFE diaphragms. INFORMATION Sterile filters must be clean and dry at all times, and should thus ideally be replaced after each use.

  • Page 62: Hoses And Accessories

    Minifors 2 - Benchtop Bioreactor Accessories Ø 25 mm, not marked Application Reagent bottles (pressure equali- sation) Retention rate 0.45 µm Diaphragm PTFE 5.17 Hoses and Accessories The following hoses and accessories, such as clamps and brack- ets, are available: Hose type Ø...

  • Page 63: O-Rings And Gaskets

    Minifors 2 - Benchtop Bioreactor Accessories 5.18 O-Rings and Gaskets Designation Ø [mm] Application Top plate gasket, culture O-ring, EPDM 3.53 x 94.84 vessel, NW90 O-ring, EPDM 3.53 x 120.24 Top plate gasket, culture vessel, NW115 Top plate gasket, culture O-ring, EPDM 3.53 x 148.8...

  • Page 64: Starter Kit

    Minifors 2 - Benchtop Bioreactor Accessories Hexagon socket spanner, WAF 17 For blanking plugs in 12 mm/Pg13.5 ports Torx wrench, TX25 For screws on the thermal block adapter 5.20 Starter Kit Each equipment package comes with a starter kit with a variety of hoses, attachments, inoculation accessories and tools.

  • Page 65: Transport And Storage

    Minifors 2 - Benchtop Bioreactor Transport and Storage 6 Transport and Storage The following specifications are based on transport and storage of an unpacked equipment at the provider’s site. 6.1 Transport WARNING Improper transport, the use of incorrect auxiliary equipment and careless handling of the equipment may lead to injuries and se- vere property damage.

  • Page 66: Storage

    Minifors 2 - Benchtop Bioreactor Transport and Storage 6.2 Storage Before each time they are put into storage, decontaminate,  thoroughly clean and dry the culture vessel and all accesso- ries Store the equipment and its components clean, dry and pro- ...

  • Page 67: Installation And Initial Operation

    Minifors 2 - Benchtop Bioreactor Installation and Initial Operation 7 Installation and Initial Operation To set up and connect the equipment, note the following: WARNING Faulty installation may lead to dangerous situations or severe loss of property. Follow the installation and commissioning instructions in this op- erating manual precisely.

  • Page 68: Power Supply

    Minifors 2 - Benchtop Bioreactor Installation and Initial Operation 7.3.1 Power Supply Connection conditions The in-house power supply to the equipment must meet the follow- ing requirements: Single-phase, constant power supply  120/230 VAC 50/60 Hz  Connection To connect the basic unit to the in-house power supply, proceed as...

  • Page 69: Gas Supply

    Minifors 2 - Benchtop Bioreactor Installation and Initial Operation Position the pressure hoses on the appropriately marked hose nozzles on the basic unit. Connect the hoses to the in-house water supply and drainage. Secure the hoses with hose clamps to prevent slipping.

  • Page 70: Exit Gas

    Minifors 2 - Benchtop Bioreactor Installation and Initial Operation WARNING The use of inappropriate or damaged hoses and/or inappropri- ate fixing may lead to leakage of gases. Depending on the gas in question, there may be a danger of gas explosion and/or dan- ger of suffocation as well as a hazard for the health of the oper- ator.

  • Page 71: Test Run

    Minifors 2 - Benchtop Bioreactor Installation and Initial Operation Insert the other plug into the socket on the motor. Depending on the size of the culture vessel, the large (left- hand) or small (right-hand) motor is supplied. 7.5 Test Run In order to become familiar with the basic functions of the equip- ment before the first cultivation, a short test run can be executed.

  • Page 72: Cooling System

    Minifors 2 - Benchtop Bioreactor Installation and Initial Operation ATTENTION If the vessel top plate presses against long components such as the stirrer shaft etc., they could bend because of the weight of the top plate. Always position the vessel top plate so that it does not lie on top of components.

  • Page 73: Stirring

    Minifors 2 - Benchtop Bioreactor Installation and Initial Operation Start the Batch (process) using Start Batch and switch on the Temperature parameter. All parameters except for Temperature remain switched off; switch them off if necessary. You should now hear water flowing into the temperature con- trol system.

  • Page 74: Gassing

    Minifors 2 - Benchtop Bioreactor Installation and Initial Operation Wait until the temperature has adjusted to the setpoint. For the rest of the procedure, allow the Batch to run with the tem- perature switched on and the stirrer running. 7.5.5...
  • Page 75 Minifors 2 - Benchtop Bioreactor Installation and Initial Operation When the motor has cooled down: Uncouple the motor from the vessel and place it on a clean and dry work surface. Empty the culture vessel. Page 73 of 214 07 February 2019...

  • Page 76: Before Cultivation

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8 Before Cultivation The following chapters describe all the preparatory work before starting the cultivation process. This essentially comprises: Preparing and autoclaving the culture vessel:  Checking the gaskets (O-rings) on component parts and ...

  • Page 77: Mounting The Impellers

    Minifors 2 - Benchtop Bioreactor Before Cultivation Procedure Check the O-ring for sealing the top plate for damage and to ensure that it is positioned correctly in the groove on the ves- sel flange. Ensure that every component part is equipped with an intact...

  • Page 78: Mounting Dip Tubes And Spargers

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.3 Mounting Dip Tubes and Spargers Straight spargers and dip tubes can be mounted to the outside of the vessel top plate. Curved spargers and dip tubes can only be mounted to the inside of the vessel top plate.

  • Page 79: Inserting The Vessel Into The Vessel Holder

    Minifors 2 - Benchtop Bioreactor Before Cultivation Set the desired mounting depth. Tighten the slotted-head screw. 8.1.4 Inserting the Vessel into the Vessel Holder To insert the glass vessel into the vessel holder, proceed as fol- lows: Procedure Place the flange onto the ring of the vessel holder: the two opposing recesses on the flange fit with the bolt on the ring.

  • Page 80: Inserting The Baffles

    Minifors 2 - Benchtop Bioreactor Before Cultivation Insert the vessel carefully. 8.1.5 Inserting the Baffles Proceed as follows: Procedure Insert the baffles carefully into the glass vessel. Page 78 of 214 07 February 2019...

  • Page 81: Moistening/Filling The Culture Vessel

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.6 Moistening/Filling the Culture Vessel If the medium in the culture vessel is to be autoclaved, this can take place before the top plate is put in position and the additional component parts are mounted.

  • Page 82: Mounting The Blanking Plugs

    Minifors 2 - Benchtop Bioreactor Before Cultivation Tighten the knurled screws on the top plate by hand (no tool!) crossways. ATTENTION If the screws are tightened too much, components may be dam- aged, which can result in failure of the equipment. The screws may not be tightened with a tool under any circumstances.

  • Page 83: Mounting Addition Port Adapters

    Minifors 2 - Benchtop Bioreactor Before Cultivation Ø 12 mm / Pg13.5 ports Procedure Insert the O-ring and blanking plug into all unused ports. Tighten by hand. Use the hexagon socket spanner to make it hand-tight. 8.1.9 Mounting Addition Port Adapters...

  • Page 84: Mounting The Feed Needle(S)

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.10 Mounting the Feed Needle(s) The procedure for mounting one or more feed needle(s) instead of addition port adapters is the same as for the mounting of the addi- tion port adapters. For details, see the chapter “Mounting Addition Port Adapters”.

  • Page 85: Preparing The Dip Tube/Addition Port Adapter For Inoculation

    Minifors 2 - Benchtop Bioreactor Before Cultivation Insert the septum (inoculation diaphragm) into the port. Screw the septum collar into the port by hand. Insert the blanking plug equipped with an O-ring into the sep- tum collar and screw it tight by hand.

  • Page 86: Mounting The Exit Gas Cooler

    Minifors 2 - Benchtop Bioreactor Before Cultivation Equip the hose for a sterile hose connection. (Depending on the application: rapid coupling, sterile connector or weldable hose with sterile filter). Secure the hose transition points with cable ties. 8.1.14 Mounting the Exit Gas Cooler...

  • Page 87: Preparing The Sensors

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.15 Preparing the Sensors All sensors that come into contact with the medium are mounted before autoclaving and are sterilised together with the culture ves- sel. Note the following about all sensors: Mount all sensors by hand – do not use any tools! ...

  • Page 88: Calibrating The Ph Sensor

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.16 Calibrating the pH Sensor For a reliable pH measurement, a 2-point calibration with an upper and lower reference buffer must be carried out before each cultiva- tion. The pH sensor must be calibrated before autoclaving. This is carried out on the operating panel.

  • Page 89: Mounting Sensors With Electrode Holder

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.18 Mounting Sensors with Electrode Holder For the mounting of a sensor in a 12 mm/Pg13.5 port for culture vessels with a nominal width of 115, an electrode holder must be used. To do so, proceed as follows:...
  • Page 90 Minifors 2 - Benchtop Bioreactor Before Cultivation Insert the sensor into the hollow screw with the thread pointing in the downward direction. Fit the fork of the guide bar into the groove of the hollow screw. Push the hollow screw and the guide bar together upwards and insert the guide bar into the hole of the support guide.

  • Page 91: Mounting The Antifoam Sensor

    Minifors 2 - Benchtop Bioreactor Before Cultivation Screw the sensor on the hollow screw into the port and tighten 10. Tighten the grub screw in the support guide with the key. 8.1.19 Mounting the Antifoam Sensor Please note the following points for mounting: The antifoam sensor is equipped with transparent insulation ...
  • Page 92 Minifors 2 - Benchtop Bioreactor Before Cultivation Fix the clamping adapter with the fastening screw. Loosen the slotted-head screw at the clamping adaptor. Set the desired mounting depth of the sensor carefully. Tighten the slotted-head screw carefully. ATTENTION If the sensor is fixed too tightly in the clamping adapter, or the...

  • Page 93: Preparing The Super Safe Sampler

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.20 Preparing the Super Safe Sampler INFORMATION The following figures are for general purposes of comprehen- sion. In order to prepare the Super Safe Sampler sampling system for autoclaving, proceed as follows: Procedure Attach the hose of the valve group on the dip tube.

  • Page 94: Mounting The Sparger Hose And The Inlet Air Filter

    Minifors 2 - Benchtop Bioreactor Before Cultivation Turn the sterile filter carefully by hand in a clockwise direction. This ensures that the non-return valve/sterile filter screw con- nection is tight. Cover the valve group loosely with aluminium foil. Clamp off the hose on the dip tube.

  • Page 95: Preparing The Reagent Bottles, Pumps And Hoses

    Minifors 2 - Benchtop Bioreactor Before Cultivation 3.0 L and 6.0 L culture vessels: filter without marking, Ø =  50 mm, fit it in either direction to the hose end. Fit the hose to the sparger. The figure to the left shows an inlet air filter for 1.5 L culture vessels as an example.
  • Page 96 Minifors 2 - Benchtop Bioreactor Before Cultivation Below is a detailed description of how reagent bottles are equipped properly and connected to the pumps and culture vessel. Connecting the reagent bottles to the pumps and culture ves- To connect the reagent bottles with the pumps and the culture ves-...
  • Page 97 Minifors 2 - Benchtop Bioreactor Before Cultivation Attach the silicone hose of the antifoam pump to the mounted antifoam sensor in the culture vessel and secure it with a ca- ble tie. Connection between reagent bottles and pumps Procedure Ensure that a hose is fitted inside the reagent bottles at the exposed hose connection (without sterile filter);...

  • Page 98: Sterile Hose Connections

    Minifors 2 - Benchtop Bioreactor Before Cultivation INFORMATION Fill reagent bottles with heat-resistant reagents only. Sterilise non-heat-resistant feed solution separately and only transfer it to the reagent bottle after sterilising. Place the reagent bottles in reagent bottle and pump holders.

  • Page 99: Setting The Pumps

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.24 Setting the Pumps If the pumps are not used with the default settings, we recommend that the appropriate settings are now made on the operating panel. It is possible, for example, to estimate and display the volume (in mL) that has been pumped since the Batch (process) started.

  • Page 100: Fitting The Cone Plug For Drive Hub

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.1.26 Fitting the Cone Plug for Drive Hub In order to prevent the penetration of condensation water into the dive hub during autoclaving, the cone plug provided in the starter set must be fitted.

  • Page 101: Autoclaving

    Minifors 2 - Benchtop Bioreactor Before Cultivation Reagent bottles are placed in reagent bottle holders and pump heads are placed on the pump holder with a mounting plate. Super Safe Sampler The valve group is connected to the dip tube in the culture vessel by means of a hose.
  • Page 102 Minifors 2 - Benchtop Bioreactor Before Cultivation INFORMATION Development of steam is not possible when autoclaving a com- pletely empty and dry culture vessel. Successful sterilisation is not guaranteed. Ensure that there is liquid in the culture vessel (approx. 10 mL of water per litre of total volume).

  • Page 103: Connecting The Culture Vessel And Preparing The Cultivation

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.2 Connecting the Culture Vessel and Preparing the Cultivation As soon as the culture vessel with the accessories has cooled suf- ficiently, it can be hung up within the basic unit and the various ca- ble and tube connections between the basic unit and the culture vessel can be established.

  • Page 104: Connecting The Gassing

    Minifors 2 - Benchtop Bioreactor Before Cultivation WARNING When using heavily corrosive reagents (acids and bases), it is particularly important only to use suitable and undamaged hoses. They must also be securely fastened. Furthermore, the exit gas filter must not be blocked. This en- sures that no pressure builds up and no reagent escapes due to burst hoses.

  • Page 105: Coupling The Motor

    Minifors 2 - Benchtop Bioreactor Before Cultivation Insert the rapid coupling of the water inlet hose – note the symbol on the basic unit - onto the lower connection nozzle on the exit gas cooler. Insert the rapid coupling of the water outlet hose – note the symbol on the basic unit - onto the upper connection nozzle on the exit gas cooler.

  • Page 106: Filling The Culture Vessel

    Minifors 2 - Benchtop Bioreactor Before Cultivation To couple the motor, proceed as follows: Procedure Place the motor onto the drive hub with the groove aligned with the pin on the drive hub. The motor is held in its position.

  • Page 107: Connecting The Temperature Sensor (Pt100)

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.2.7 Connecting the Temperature Sensor (Pt100) The temperature sensor is not in direct contact with the medium. Procedure Insert the sensor into the pocket in the vessel top plate as far as it will go..

  • Page 108: Connecting The Ph Sensor

    Minifors 2 - Benchtop Bioreactor Before Cultivation Insert the black banana plug into the earth connection in the top plate. 8.2.9 Connecting the pH Sensor To connect the pH sensor, proceed as follows: Procedure Remove the red protective cap from the sensor cable.

  • Page 109: Calibrating The Po Sensor

    Minifors 2 - Benchtop Bioreactor Before Cultivation 8.2.11 Calibrating the pO Sensor Generally speaking, the following applies: The pO sensor should be calibrated after autoclaving has been performed because the sterilisation process may change the steepness of the pO sensor.

  • Page 110: Cultivation

    Minifors 2 - Benchtop Bioreactor Cultivation 9 Cultivation The following sections describe the work necessary for the perfor- mance of and after the completion of a cultivation, before the cul- ture vessel with accessories is thoroughly cleaned and then pre- pared for another cultivation.

  • Page 111: Sampling

    Minifors 2 - Benchtop Bioreactor Cultivation For details on operation, see the corresponding chapter in the main chapter “Operation”. INFORMATION Depending on the specifications defined by the user, the pO sensor is calibrated either before the vessel is filled with medium or afterwards, in the prepared medium.
  • Page 112 Minifors 2 - Benchtop Bioreactor Cultivation WARNING Loose screws at components could lead to the penetration of unsterile air or contamination of the environment. Before and after autoclaving: Check that all screws are tightly screwed in and, if necessary, tighten them manually If the sample is to be further aseptically processed, use a sterile syringe and sterile closing caps.
  • Page 113 Minifors 2 - Benchtop Bioreactor Cultivation If the dip tube was rinsed with air, air is sucked in first. Remove it as follows: Unscrew the syringe from the valve. Hold the syringe with the plunger downwards so that the me- dium remains in the syringe.

  • Page 114: Inoculation

    Minifors 2 - Benchtop Bioreactor Cultivation Procedure: Hold the syringe with sample downwards, pull back the plunger. This removes all but a few µL of the residual fluid. Hold the sample valve with one hand; unscrew the syringe with the other.

  • Page 115: Inoculation With A Syringe

    Minifors 2 - Benchtop Bioreactor Cultivation All clamps have been removed (except for sampling system).  Utensils for the inoculation and vessels with inoculum are  ready. Methods There are a number of ways to add medium or inoculum before...

  • Page 116: Harvest

    Minifors 2 - Benchtop Bioreactor Cultivation 9.4 Harvest The culture can be harvested at the end of the cultivation. To pre- vent possible sedimentation from the culture, the stirrer can be switched on during harvesting. If necessary, activate gassing for sensitive cultures.

  • Page 117: Emptying The Culture Vessel

    Minifors 2 - Benchtop Bioreactor Cultivation 9.5 Emptying the Culture Vessel Depending on the user specifications, the culture vessel can be emptied either before or after autoclaving. A previously emptied and culture vessel filled only with water for autoclaving is easier to clean afterwards.

  • Page 118: Autoclaving The Culture Vessel After Cultivation

    Minifors 2 - Benchtop Bioreactor Cultivation INFORMATION Always stop the running Batch (process) on the operating panel. If it is stopped by pressing the main switch, it is akin to a power interruption. This means that when it is switched on again, the...
  • Page 119 Minifors 2 - Benchtop Bioreactor Cultivation Remove all cable and hose connections between the basic unit and the culture vessel: Uncouple the motor and place it to the side. Unplug the sensor cables. Pull the temperature sensor out of the pocket.

  • Page 120: Operation

    Minifors 2 - Benchtop Bioreactor Operation 10 Operation 10.1 Screen Areas, Menu Navigation and Control Elements Left side with selection menus for system set- Buttons for the selection of the parameter dis- tings or parameter groups play and time stamp for sampling...
  • Page 121 Minifors 2 - Benchtop Bioreactor Operation Display of the current time  Display for connected USB stick  Display for an active connection to SCADA software  Display for an active wireless connection  Alarm display If alarms occur (equipment alarm or parameter alarm), they are signalled by a red exclamation mark highlighted in white on a red background.

  • Page 122: Main Screen

    Minifors 2 - Benchtop Bioreactor Operation 10.1.1 Main Screen Depending on the menu selected on the left side of the screen, the main screen displays the following: Menus for system settings, such as the VESSEL TYPE menu for setting the vessel size.

  • Page 123: Edit View

    Minifors 2 - Benchtop Bioreactor Operation All menus and parameter groups can be selected by pressing them. The selected menu or parameter group is highlighted with a change of colour in the menu/group text from grey to orange. Example to the left: MAIN parameter group The arrow buttons at the edge of the main screen can be used to show or hide parts of the menu and display.

  • Page 124: Start Batch / Inoculate / Stop Batch

    Minifors 2 - Benchtop Bioreactor Operation OK confirms the selection and closes the menu.  CANCEL closes the menu without making any changes.  10.1.3 START BATCH / INOCULATE / STOP BATCH Pressing the START BATCH button starts the preparation phase for the Batch (bioprocess).

  • Page 125: Sample Now

    Minifors 2 - Benchtop Bioreactor Operation After pressing STOP BATCH, the Stop Process dialogue appears asking for confirmation that the Batch is to be stopped, as well as the notice that doing so deactivates all parameters. CANCEL cancels the stop procedure without making any ...

  • Page 126: Menus For System Settings

    Minifors 2 - Benchtop Bioreactor Operation This means that the Sample information dialogue appears and dis- plays how long the batch has been running since inoculation (Batch Time….), and how many samples have been taken (Sam- ple Nr.….), or how many times SAMPLE NOW has been pressed since START BATCH was pressed.

  • Page 127: Vessel Type - Selecting A Culture Vessel

    Minifors 2 - Benchtop Bioreactor Operation INFORMATION Depending on the menu selected, the CANCEL (leave the menu without changes) and OK (save changes and leave the menu) buttons, or only the OK button, are available in the menu footer. 10.2.1 VESSEL TYPE – Selecting a Culture Vessel The culture vessel used is set in the VESSEL TYPE menu.

  • Page 128: Appearance - Display Settings

    The languages in the pull-down menu are always displayed in English. INFORMATION Additional languages can be downloaded from the INFORS HT website to a USB stick and loaded to the equipment from the USB menu. If the desired language is not available, contact the local service partner of the equipment manufacturer.
  • Page 129 Minifors 2 - Benchtop Bioreactor Operation DISPLAY INFO BUTTONS Switching the on-screen help off or on This display of the info buttons for the on-screen help for the vari- ous parameters is switched on (YES) or off (NO) using the switch.

  • Page 130: Network Settings

    Minifors 2 - Benchtop Bioreactor Operation ACCEPT DATE AND TIME FROM OPC Switching on or off the acceptance of date and time from a PC If this function is switched on, a connected OPC UA client such as the bioprocess platform software eve can overwrite the date and ®...
  • Page 131 Minifors 2 - Benchtop Bioreactor Operation CONNECTION TYPE Select whether a cable connection (Ethernet) or cable-free (wire- less) network connection is to be used. An optional wireless adaptor is required for a connection with a wireless network. CONFIGURATION Determine whether the network connection is to be automatically configured (Auto (DHCP)) or needs to be set up manually (Man- ual).

  • Page 132: Eve Communication - Communication Settings

    Minifors 2 - Benchtop Bioreactor Operation 10.2.4 eve COMMUNICATION – Communication Settings In the eve COMMUNICATION menu, permissions for server ac- cess as well as their security settings for communication with the bioprocess platform software eve are set. ® INFORMATION Display of the equipment name (Device Name) and its network ad- dress (IP, configuration under NETWORK SETTINGS).

  • Page 133: Usb Data Export And Import From A Usb Stick

    Minifors 2 - Benchtop Bioreactor Operation 10.2.5 USB Data Export and Import from a USB Stick In the USB menu, a USB stick connected with the equipment’s USB port can be used to import or export data. EXPORT DATA TO USB Opens the menu for data export.
  • Page 134 Minifors 2 - Benchtop Bioreactor Operation Four files are created per batch and are ready for export. Each file name contains the start date and start time of the batch: yyyy Year Month Hours Minutes Seconds The four files contain the following: yyyy_mm_dd_hh_ii_ss_Parameters.json...
  • Page 135 UPDATE USER INTERFACE A selection menu for data import appears. An update for the oper- ating panel can be selected, e.g. downloaded from the INFORS HT download centre or provided by an authorised INFORS HT service partner.

  • Page 136: System Info - System Information

    Minifors 2 - Benchtop Bioreactor Operation 10.2.6 SYSTEM INFO – System Information In the SYSTEM INFO menu, the currently installed version of the firmware and the operating hours since start-up are displayed. 10.3 Parameter - Parameter Groups On the main screen, up to eight parameters can be simultaneously monitored and controlled.

  • Page 137: Parameters - Displays And Functions

    Minifors 2 - Benchtop Bioreactor Operation FAVOURITES: Allows you to compile up to eight parameters  from the other four parameter groups. This can be done using the EDIT VIEW button. For details, see the “EDIT VIEW” sec- tion. MAIN: Contains the Temperature, Stirrer, pH, pO , TotalFlow, ...
  • Page 138 Minifors 2 - Benchtop Bioreactor Operation ON/OFF switches control of the selected parameter on or off INFORMATION ON/OFF is only available when a Batch is running. First start the batch with START BATCH and, if necessary, INOCULATE. Calibrating opens the calibration menu of the selected parameter.

  • Page 139: Setpoint - Setting The Setpoint

    Minifors 2 - Benchtop Bioreactor Operation 10.3.2 SETPOINT - Setting the Setpoint The setpoints can be entered in any operating state of the equip- ment for parameters that are not controlled via cascade and have a controller output. Parameter control is however only active when a batch has been started using START BATCH and the correspond- ing parameter has been activated using ON/OFF.

  • Page 140: Parameter Alarms

    Minifors 2 - Benchtop Bioreactor Operation Example: The Stirrer setpoint entry is too high. Make a new entry within the permissible values of 0 to 1,600 min 10.3.3 Parameter Alarms If a parameter is activated and the Batch inoculated, parameter alarms are generated after a predefined waiting time if there are unexpected deviations from the actual values and setpoints.
  • Page 141 Minifors 2 - Benchtop Bioreactor Operation In the parameter group that contains the parameter in ques- tion, a number on a red background appears. This indicates the number of existing parameter alarms. The parameter in question is displayed with a red bar and a red actual value.

  • Page 142: Cascades

    Minifors 2 - Benchtop Bioreactor Operation 10.3.4 Cascades Cascades can be configured for some parameters. A cascade can be used to assign a parameter to another parameter as an actua- tor. Example: For control of the pO by changing the Gasmix parameter, a cas- cade to the Gasmix parameter is configured for the pO .

  • Page 143: Main Parameter Group

    Minifors 2 - Benchtop Bioreactor Operation 10.4 MAIN Parameter Group The MAIN parameter group contains all parameters that are availa- ble as standard, as well as the GasMix and TotalFlow parameters, which are only functional when using a second (optional) gas.
  • Page 144 Minifors 2 - Benchtop Bioreactor Operation For details on the safety, technical data, usage and maintenance requirements for the pH sensors, see the separate documentation provided by the sensor manufacturer. The pH control can be configured using a cascade and takes place by default by adding acid and base via the two peristaltic pumps Pump1/Acid and Pump2/Base.

  • Page 145: Po 2

    Minifors 2 - Benchtop Bioreactor Operation The PID settings can be adjusted here as required or, if necessary, can be reset to factory settings using RESET PID. For details on the PID controller, see the “PID Controller - Basics” section and associated sub-sections.
  • Page 146 Minifors 2 - Benchtop Bioreactor Operation At the CASCADE menu item, the pull-down menu is used to call up the list of pre-defined cascades for pO control. The following settings are available for selection: None: No control, pO is only measured.
  • Page 147 Minifors 2 - Benchtop Bioreactor Operation The PID settings can be adjusted here as required or, if necessary, can be reset to factory settings using RESET PID. For details on the PID controller, see the chapter “PID Controller - Basics” and associated subchapters.

  • Page 148: Totalflow

    Minifors 2 - Benchtop Bioreactor Operation 10.4.5 TotalFlow Measures and controls the sum of the volume flows of air (AirFlow) and the (optional) second connected gas (Gas2Flow). The mixing ratio of air with a second, optional gas is controlled by the GasMix parameter.

  • Page 149: Foam

    Minifors 2 - Benchtop Bioreactor Operation Only Air: Air is exclusively used, with no addition of a second  gas. TotalFlow therefore remains constant and corresponds to AirFlow. Air/O2: If the setpoint is greater than 21 %, O is additionally ...
  • Page 150 Minifors 2 - Benchtop Bioreactor Operation AntiFoam: Addition of antifoaming agent upon detection of  foam Level: Pump-down of culture medium when a certain fill level  is reached If the Antifoam or Level function is selected, additional parameter settings are possible: DOSE TIME: Duration (in seconds) of the addition of anti- ...

  • Page 151: Extended Parameter Group

    9600, 8 bits, no parity, 2 stop bits. For a list of compatible balances or help with the connection, please contact your local INFORS HT service partner. 10.5.3 AirFlow Measures and controls the flow rate of air into the culture vessel...

  • Page 152: Gas2Flow

    Minifors 2 - Benchtop Bioreactor Operation valve). The measurement system is completely electronic and the measurement value is displayed in L/min.) INFORMATION The mass flow controller is calibrated by the manufacturer ex works at standard conditions, i.e. at 1.013 bar and 20 °C. There-...

  • Page 153: Exitgas Parameter Group

    Minifors 2 - Benchtop Bioreactor Operation INFORMATION If TotalFlow and Gasmix are used to control gassing, it is not possible to individually enter setpoints for Gas2Flow. Gas2Flow can only be controlled individually, when TotalFlow and Gasmix are deactivated. 10.6 EXITGAS Parameter Group The EXITGAS parameter group contains the parameters for the optional exit gas analysis.

  • Page 154: Exit Gas Co

    Minifors 2 - Benchtop Bioreactor Operation 10.6.2 Exit Gas CO Measures the carbon dioxide concentration in the exit gas of the bioreactor using a combined BlueInOne Ferm gas sensor (see Exit Gas CO Measurement range depends on the chosen variant of the gas sensor: 0 to 10 Vol.
  • Page 155 Minifors 2 - Benchtop Bioreactor Operation Example Analogue: 50 % = half speed = half delivery capacity  Digital: 50 % = 100 % speed, but only active 50 % of the time  = half delivery rate. Ex works, the pumps are configured as follows: Pump 1: ACID (addition of acid, digital, with fixed speed), ...

  • Page 156: Configuring The Pumps

    Minifors 2 - Benchtop Bioreactor Operation 10.7.1 Configuring the Pumps The editor menu of every pump has four menu items for configura- tion. The figure below shows the editor menu of Pump 1 as an ex- ample. TUBE TYPE Select the pump hose used.

  • Page 157: Pump1 - Acid Or Additional Feed Solution

    Minifors 2 - Benchtop Bioreactor Operation FEATURE Sets the pump function mode Since the four pumps have different function modes, they are de- scribed in the following sub-sections. DISPLAY COUNT UNIT Configures the display of the totalled actual value. Select either Count (number of rotations of the pump head) or ~ml (the pumped volume estimated on the basis of the hose diameter selected under TUBE TYPE).

  • Page 158: Pump2 - Base Or Additional Feed Solution

    Minifors 2 - Benchtop Bioreactor Operation INFORMATION The function mode of Pump1 can also be changed by making corresponding entries in the editor menu of the pH parameter. 10.7.3 Pump2 - Base or Additional Feed Solution Pump2 can be configured for the Base operating mode (factory setting) or Feed.

  • Page 159: Pump4 - Feed Solution

    Minifors 2 - Benchtop Bioreactor Operation INFORMATION The function mode of Pump3 can also be changed by making corresponding entries in the editor menu of the Foam parame- ter. 10.7.5 Pump4 - Feed Solution Pump4 can be configured for the Feed function mode (factory set- ting) or, provided an optional balance is connected and the Bal- ance parameter is available, it can be configured for Balance Feed.

  • Page 160: Auto Fill/Empty - Automatically Filling/Emptying Pump Hoses

    Minifors 2 - Benchtop Bioreactor Operation 10.7.6 AUTO FILL/EMPTY – Automatically Filling/Emptying Pump Hoses Using OPEN AUTO FILL/EMPTY in the pump menu, the sub-menu for automatic filling and emptying of the pump hoses is opened. For each pump, an individual filling duration (FILLING DURATION) as well as an individual emptying duration ( EMPTYING DURA- TION) can be defined.

  • Page 161: Calibration

    Minifors 2 - Benchtop Bioreactor Operation If a filling or emptying procedure is active, the remaining filling or emptying time is displayed. In the example to the left, the remaining filling time is displayed. The menu cannot be closed while at least one filling or emptying procedure is active.

  • Page 162: Calibrating The Ph Sensor - Procedure

    Minifors 2 - Benchtop Bioreactor Operation The pHs and the temperature dependence of pH buffers are saved in the pH sensors and are automatically detected during calibra- tion. It is therefore not necessary to carry out a separate tempera- ture measurement of the buffer solution.
  • Page 163 Minifors 2 - Benchtop Bioreactor Operation Header: Date of the last calibration  2-POINT CALIBRATION: Select 2-point calibration  PRODUCT CALIBRATION: Select product calibration (for  details, see the “pH product calibration” section). SHOW SENSOR STATUS: Shows the data and values ...
  • Page 164 Minifors 2 - Benchtop Bioreactor Operation 3 Wait until measurement is stable: Wait until the meas-  ured value is stable. 4 Perform the calibration at the first point: Start the cali-  bration procedure for the first calibration point.
  • Page 165 Minifors 2 - Benchtop Bioreactor Operation In the pull-down menu, either select the Auto entry or the ap- propriate value of the possible pH reference value. INFORMATION The pH sensors are calibrated with the following standard set- tings ex works: Buffer pH 4 (calibration point 1) buffer pH 7 (cali- bration point 2) at room temperature.
  • Page 166 Minifors 2 - Benchtop Bioreactor Operation Rinse the pH sensor with distilled water. Do not rub it. Hold the pH sensor in the buffer solution of the second calibration point, e.g. a solution of pH 7.00. 10. In the pull-down menu, either select the Auto entry or the ap- propriate value of the possible pH reference value.

  • Page 167: Ph Sensor Product Calibration

    Minifors 2 - Benchtop Bioreactor Operation 10.8.3 pH Sensor Product Calibration It is possible to adjust the calibration curve to the current process conditions using product calibration. This could be necessary if there is a possibility of drift of the displayed pH during a long-term cultivation, for example.
  • Page 168 Minifors 2 - Benchtop Bioreactor Operation Left-hand side 1 Take sample for offline measurement and confirm:  Take a sample for external measurement and confirm. Sample was taken at: Display of date and time of the  sampling. 2 Measure the pH of the sample and enter the value: ...
  • Page 169 Minifors 2 - Benchtop Bioreactor Operation Take a sample from the process (in the culture vessel). There are two possible approaches: Confirm the sampling (generate a time stamp), carry out a laboratory measurement of the pH value for the sam- ple, enter the measured value and carry out product cali- bration.

  • Page 170: Calibrating The Po Information

    Minifors 2 - Benchtop Bioreactor Operation In the calibration menu, Active is displayed under PRODUCT CALIBRATION to show that a product calibration was carried out and is active. INFORMATION The original calibration curve can be restored again using a 2- point calibration.

  • Page 171: Calibrating The Po Sensor - Procedure

    Minifors 2 - Benchtop Bioreactor Operation The zero point (0 % calibration) should be checked at regular inter- vals of approximately 6 months. When carrying out a 2-point calibration, the 0 % calibration must be carried out before the 100 % calibration.
  • Page 172 Minifors 2 - Benchtop Bioreactor Operation Proceed as follows: Procedure Connect the nitrogen to the gassing line (O IN connec- tion). In the process, leave the gas supply closed. INFORMATION If necessary, enter setpoints for temperature and pH, activate parameters and press START BATCH and wait until the desired operating temperature and the expected pH have been reached.
  • Page 173 Minifors 2 - Benchtop Bioreactor Operation After the initialisation phase, the menu display changes: Header: Date of the last calibration  1 POINT CALIBRATION: Select 1-point calibration  2 POINT CALIBRATION: Select 2-point calibration  SHOW SENSOR STATUS: Shows data and values ...
  • Page 174 Minifors 2 - Benchtop Bioreactor Operation The menu display changes and displays the following: Left-hand side First Calibration: First calibration point  1 Select the value of first calibration point: Select the  reference value of the first calibration point 2 Optionally set setpoints for XX % pO2: If necessary, ...
  • Page 175 Minifors 2 - Benchtop Bioreactor Operation Right-hand side Pull-down-menu for the selection of 0 % or 100 % calibration  point. Input field for the desired pO setpoint in % (depending on  the input, activates the corresponding gassing with air/nitro-...
  • Page 176 Minifors 2 - Benchtop Bioreactor Operation Enter the setpoint 0 (%) in the input field. The input field turns orange and displays the set setpoint. A dialogue appears with the notice Make sure that N2 is con- nected to the Gas2 connector Make sure that N2 is connected to the correct connection.
  • Page 177 Minifors 2 - Benchtop Bioreactor Operation If the calibration was successful, the menu display changes automatically to calibrate the second point. 10. In the pull-down-menu, select the value 100 (%). The selection is displayed. CALIBRATE POINT 2 (or 1, if not calibrated one after the other in 1-point calibration mode) is activated, i.e.

  • Page 178: Calibrating The Turbidity Sensor - General Information

    Minifors 2 - Benchtop Bioreactor Operation 12. Wait until the medium is saturated with oxygen, i.e. wait until the measured value (display via CALIBRATE POINT 2) is sta- ble. 13. Press CALIBRATE POINT 2 (or 1) to begin calibration. Gassing and the stirrer are stopped.
  • Page 179 Minifors 2 - Benchtop Bioreactor Operation In the header, the menu displays the date of the last calibra- tion, and the calibration sequence appears to the left: 1 Wait until measurement is stable: Wait until the measured  value is stable.

  • Page 180: Pid Controller - Basic Principle

    Minifors 2 - Benchtop Bioreactor Operation 10.9 PID Controller – Basic Principle PID controllers (Proportional Integral Derivative controller) are used for some of the parameters. The PID function is based on a generic formula provided as example: Explanation of the formula Error = deviation between setpoint value and actual value.
  • Page 181 Minifors 2 - Benchtop Bioreactor Operation The ideal approach is to switch off the equipment briefly after changing the integral factor in order to delete the pending error cal- culation. A typical integral factor is < 0.05. It should equate to the reciprocal value of double to quadruple the system’s cycle duration.

  • Page 182: Useful Information For Changing Pid Controller Settings

    Minifors 2 - Benchtop Bioreactor Operation 10.9.1 Table with Setting Values for PID Controller Setting value Description P (Prop. Term) Proportional factor: The greater the discrepancy between the set- point value and the actual value the greater the controller output I (Integ.

  • Page 183: Adjusting Pid Settings

    Also see the “Parameter alarms” section. Equipment errors: If equipment errors occur repeatedly or  cannot be resolved, inform an authorised INFORS HT service partner. Also see the fault table in the “Interferences” section. The Equipment Alarm menu is only available when there are open or unconfirmed alarms.
  • Page 184 Minifors 2 - Benchtop Bioreactor Operation Pressing the alarm symbol or swiping upwards opens the Equip- ment Alarm menu. DESCRIPTION: Display of the type of alarm.  STATE: Status display of the alarm, open or resolved  Open alarms are displayed in red and with the word ...

  • Page 185: Cleaning And Maintenance

    Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance 11 Cleaning and Maintenance The following sections describe in detail how the culture vessel and accessories and the basic unit are cleaned and, as required, stored. In addition, the section contains a maintenance plan and corre- sponding descriptions for the procedures to be performed by the operator.
  • Page 186 Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance The following method describes a routine cleaning between two fermentations/cultivations. It takes place with the culture vessel completely assembled and the accessories completely mounted. This does not include the sensors, with the exception of antifoam or level sensors from the equipment manufacturer.

  • Page 187: Removing The Vessel Top Plate And Accessories

    Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance 11.3 Removing the Vessel Top Plate and Accessories All accessories must be removed for thorough cleaning of the indi- vidual parts of the culture vessel. This is described in the following sections. The cleaning itself is described in the chapter “Cleaning and Storing Individual Parts”.
  • Page 188 Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance INFORMATION The sensor can be pulled out of the vessel top plate port along with the clamping adapter. After subsequently unscrewing the slotted-head screw on the clamping adapter, the sensor can be pulled out of the clamping adapter.
  • Page 189 Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance 11.3.4 Removing Blanking Plugs Proceed as follows: Blanking plugs in 10 mm vessel top plate ports Procedure Loosen and remove the fastening screw beside the blanking plugs by hand. Pull the blanking plug out of the vessel top plate port by hand.
  • Page 190 Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance Procedure As far as possible, remove mounted parts before lifting the top plate. Remove the knurled screws on the top plate by hand (no tool!) and place them to the side. Carefully lift the top plate vertically upwards from the vessel until the stirrer shaft and other long components can no longer come into contact with the glass vessel.

  • Page 191: Cleaning And Storing Individual Parts

    Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance 11.3.9 Removing the Impellers Before removing the impellers, it is recommended to measure and record the positions to aid later mounting. Proceed as follows to remove: Loosen the grub screws on the impeller with an Allen key – do...

  • Page 192: Cleaning The Sensors

    Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance Procedure Clean parts with distilled water and a soft sponge or in the dishwasher. Ensure that the deposits in the dip tubes and, if necessary, the feed needles, are removed. Use 0.1 N caustic soda solution followed by distilled water as necessary.

  • Page 193: Cleaning The Super Safe Sampler

    Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance 11.7 Cleaning the Super Safe Sampler ATTENTION Risk of damage to the sampling system from unsuitable clean- ing methods or cleaning agent (such as acids, bases or sol- vents, for example). –...

  • Page 194: Cleaning The Basic Unit And Operating Panel

    Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance Flush the exit gas cooler with ethanol (70%). Thoroughly rinse the exit gas cooler with distilled water. 11.9 Cleaning the Basic Unit and Operating Panel Proceed as follows to clean the surface of the basic unit and the...

  • Page 195: Decalcifying The Equipment

    Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance To be carried out by operator Interval Maintenance work Before each cultivation Check all hoses and hose lines. Check cables for damage and kinks. Check that O-rings and gaskets are leak-proof, replace if necessary.
  • Page 196 Minifors 2 - Benchtop Bioreactor Cleaning and Maintenance Ensure that the valve for the exit gas cooler water supply is open. Open it, if necessary. Hang the culture vessel on the basic unit (hang the vessel holder on hooks on the thermal block).

  • Page 197: Interferences

    Minifors 2 - Benchtop Bioreactor Interferences 12 Interferences The following section describes possible reasons for interferences and how to resolve them. Reduce the service intervals in corre- spondence with the actual loads if interferences become increas- ingly common. Contact the manufacturer for interferences that can- not be resolved by following the above instructions.

  • Page 198: Interferences Drive System

    Minifors 2 - Benchtop Bioreactor Interferences 12.2 Interferences Drive System Interference Motor does not start. Possible cause Remedy Check cable connections and connect correctly as Motor not properly connected. Operator necessary. The Stirrer parameter is not acti- Activate the Stirrer parameter.

  • Page 199: Interferences Temperature Control System

    Minifors 2 - Benchtop Bioreactor Interferences 12.3 Interferences Temperature Control System Interferences No temperature control. Possible cause Remedy Parameter Temperature is not acti- Activate the parameter. Operator vated. Parameter Stirrer is not activated. Activate the parameter. Operator Interference No cooling or inadequate cooling.

  • Page 200: Interferences Ph-System

    Minifors 2 - Benchtop Bioreactor Interferences Exit gas filter blocked. The overpressure sensor switches gassing off for 10 Operator sec, replace the exit gas filter under sterile condi- tions. Interference The desired gas flow rate is not reached. Possible cause Remedy Blocked holes on the sparger.
  • Page 201 Minifors 2 - Benchtop Bioreactor Interferences Interference No pH control. Possible cause Remedy The pH parameter is not activated. Activate the pH parameter. Operator Pumps are not switched on. Switch on pump1 (Acid), pump2 (Base) Operator Incorrect dead band setting.

  • Page 202: Interferences Po

    Minifors 2 - Benchtop Bioreactor Interferences 12.6 Interferences pO System Interferences No display or incorrect display of pO Possible cause Remedy Sensor cable not connected or not Check the sensor cable, connect it properly if neces- Operator properly connected. sary.

  • Page 203: Interferences Antifoam/Level Sensor And Antifoam

    Operator sary. Interference Foam/medium is always/frequently detected. Possible cause Remedy Sheathing of antifoam sensor is Have the sheathing of the antifoam sensor replaced. INFORS HT damaged. service techni- cian or li- censed dealer Interference Antifoam pump does not work. Possible cause...

  • Page 204: Interferences Addition Of Feed Solution (Feed Pump)

    Minifors 2 - Benchtop Bioreactor Interferences 12.8 Interferences Addition of Feed Solution (Feed Pump) Interference No feed solution or inadequate feed solution. Possible cause Remedy The Feed parameter (pump) is not Activate the Feed parameter (pump). Operator activated. Feed parameter (pump) setpoint = 0. Set Feed parameter (pump) setpoint > 0.

  • Page 205: Disassembly And Disposal

    Minifors 2 - Benchtop Bioreactor Disassembly and Disposal 13 Disassembly and Disposal The equipment must be disassembled and disposed of in an envi- ronmentally friendly manner if it is no longer in use. INFORMATION When returning the equipment for disassembly or disposal, it is...
  • Page 206 Minifors 2 - Benchtop Bioreactor Disassembly and Disposal WARNING Electronic waste, electronic components, lubricants or other auxiliary materials/supplies are subject to hazardous waste reg- ulations and may only be disposed of by registered specialist disposal firms. For disposal, the system units are to be disassembled and disman- tled into individual material groups.

  • Page 207: Technical Data

    Minifors 2 - Benchtop Bioreactor Technical Data 14 Technical Data 14.1 Equipment Dimensions Front view Top view Dimensions in mm Page 205 of 214 07 February 2019...

  • Page 208: Dimensions Of Culture Vessel

    Minifors 2 - Benchtop Bioreactor Technical Data 14.2 Dimensions of Culture Vessel The two dimension drawings show a fully equipped culture vessel ready for autoclaving. Dimensions in mm Page 206 of 214 07 February 2019...

  • Page 209: Weights (Netto)

    Minifors 2 - Benchtop Bioreactor Technical Data 14.3 Weights (netto) Basic unit Culture vessel NW 90 NW 115 NW 145 23.5 ± 0.5 kg 6 ± 0.5 kg 7.5 ± 0.5 kg 9.5 ± 0.5 kg ) Equipped culture vessel, without medium, with vessel holder. The ac- tual weight depends on design and allocation.

  • Page 210: Gas (Air, O 2 Or N 2 )

    Minifors 2 - Benchtop Bioreactor Technical Data 14.4.4 Gas (Air, O or N Description Value Unit Constant connection pressure 2 ± 0.5 Hose nozzle connection, nominal width General gas quality Dry, clean and free of oil and dust Recommended compressed air...

  • Page 211: Stirrer

    Minifors 2 - Benchtop Bioreactor Technical Data INFORMATION The volume markings on the glass vessels are only intended as visual aids. They do not represent precise measurements in li- tres. Ports Port Quantity acc. to vessel size Ø Thread NW 90...

  • Page 212: Temperature

    Minifors 2 - Benchtop Bioreactor Technical Data Accuracy Measuring: ± 5 min at 100 min to 1,600 min 1 % setpoint at > 100 % min-1 Control: 1 % full scale Direction of stirrer Anti-clockwise = to left shaft’s rotation...

  • Page 213: Gassing

    Minifors 2 - Benchtop Bioreactor Technical Data 14.5.5 Gassing All gas enters via the sparger. The specific gas flow rate, calculated for the maximum working vol- ume, is 2 min for every vessel size. Variants Gas(ses) Controlling flow rate MFC accuracy 2 MFC ±...

  • Page 214: Antifoam

    Minifors 2 - Benchtop Bioreactor Technical Data For details on the technical data, usage and maintenance require- ments for the pH sensors, see the separate documentation pro- vided by the sensor manufacturer. 14.5.7 pO Control system Via cascade Stirrer Gas flow...

  • Page 215: Pumps

    Minifors 2 - Benchtop Bioreactor Technical Data 14.5.9 Pumps Type Quantity Peristaltic Configuration Standard Alternative setting FEED, (analogue) Pump 1 ACID, digital Pump 2 Base, (digital) FEED, (analogue) Pump 3 AF = (antifoam), digi- LEVEL, (digital) FEED, (analogue) Pump 4...

  • Page 216: Auxiliary Supplies

    Minifors 2 - Benchtop Bioreactor Technical Data 14.8 Auxiliary Supplies pH buffers Intended use pH 4.0 For calibrating the pH sensor pH 7.0 Page 214 of 214 07 February 2019...

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