Infors HT Multifors 2 Operating Manual
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Infors HT Multifors 2 Operating Manual

Parallel bench-top bioreactor
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Operating Manual
2018-03-28
Multifors 2
Parallel Bench-top Bioreactor
Dok-ID: V. 04.01 - Original

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Summary of Contents for Infors HT Multifors 2

  • Page 1 Operating Manual 2018-03-28 Multifors 2 Parallel Bench-top Bioreactor Dok-ID: V. 04.01 - 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

    Multifors 2 - Parallel Bench-top Bioreactor Table of Contents General Information ............... 8 About this Manual ............8 Symbols ................. 9 1.2.1 Safety Instructions .......... 9 1.2.2 Hints and Recommendations ......9 1.2.3 Specific Safety Instructions ......10 Limitation of Liability ............ 11 Copyright Protection ............
  • Page 4 Multifors 2 - Parallel Bench-top Bioreactor Table of Contents ..............39 3.5.10 3.5.11 Pumps ............40 3.5.12 Sterile Filters ..........41 Operating Conditions ...........42 Emissions ..............42 Identification Plate ............43 Setup and Function ..............44 Basic Unit ..............44 4.1.1 Main Switch ..........46 4.1.2 Mains Connection .........46 4.1.3...
  • Page 5 Multifors 2 - Parallel Bench-top Bioreactor Table of Contents 4.7.3 Sampling System ......... 81 4.7.3.1 Super Safe Sampler, aseptic Sampling System ......81 4.7.4 Starter-Kit ............. 86 Field of Application ............87 Types ................87 Transport, Packaging and Storage ........88 Safety Instructions for Transport ........
  • Page 6 Multifors 2 - Parallel Bench-top Bioreactor Table of Contents 7.2.2.2 Mounting the pH and pO Sensors ........121 7.2.2.3 Mounting the Antifoam Sensor ... 125 7.2.3 Preparing the Reagent Bottles, Hoses and Pumps ............128 7.2.4 Calibrating the Pumps ........128 7.2.5 Connecting the Reagent Bottles to the Pumps and Vessels ..........129...
  • Page 7 Safety ................. 174 Environmental Protection ........... 176 Maintenance Plan ............176 Sterilising the Vessel after Fermentation ....178 Multifors 2 Cell: Exchanging the Ball Bearings of the Stirrer Shaft ..............180 9.5.1 Dismantling the used Ball Bearings ... 181 9.5.2 Mounting the new Ball Bearings ....

  • Page 8: General Information

    Multifors 2 - Parallel Bench-top Bioreactor General Information 1 General Information 1.1 About this Manual This manual enables the safe and efficient handling of the equip- ment. This manual is a component part of the equipment. It must be kept near to the equipment and must be accessible to the operators at all times.

  • Page 9: Symbols

    Multifors 2 - Parallel Bench-top Bioreactor General Information 1.2 Symbols 1.2.1 Safety Instructions Safety instructions are labelled using symbols. All safety instruc- tions begin with a word that signifies the degree of hazard. Strictly follow all safety points and act with due caution to avoid ac- cidents, damage to equipment, personal injuries and loss of prop- erty.

  • Page 10: Specific Safety Instructions

    Multifors 2 - Parallel Bench-top Bioreactor General Information 1.2.3 Specific Safety Instructions The following safety instructions are used to call attention to partic- ular hazards. DANGER! Danger of fatal electric shock! …signifies danger by electric current. Non-ob- servance of safety instructions may lead to a se- vere or fatal physical injury.

  • Page 11: Limitation Of Liability

    Multifors 2 - Parallel Bench-top Bioreactor General Information 1.3 Limitation of Liability All information and instructions in this manual comply with current standards and regulations, as well as the current state of technol- ogy & the manufacturer’s knowledge and experience.

  • Page 12: Spare Parts

    Multifors 2 - Parallel Bench-top Bioreactor General Information 1.5 Spare Parts WARNING! Safety risk due to incorrect spare parts! Inappropriate or faulty spare parts may impair safety and/or may lead to damage, malfunction or complete breakdown Therefore: – Use only original spare parts from the manufac- turer.

  • Page 13: Safety

    Multifors 2 - Parallel Bench-top Bioreactor Safety 2 Safety This section outlines all important safety aspects for optimal per- sonnel protection and for the safe and error-free operation of the equipment. Non-observance of the operational descriptions and safety instruc- tions listed in the operating manual may lead to serious hazards.

  • Page 14: Requirements For Qualified Personnel

    Multifors 2 - Parallel Bench-top Bioreactor Safety 2.2 Requirements for Qualified Personnel WARNING! Risk of injury when used by anyone inade- quately qualified! Inappropriate use of the equipment may lead to se- rious physical injury and material damage. Therefore: – All operations must be executed by qualified personnel only.

  • Page 15: Unauthorised Persons

    Multifors 2 - Parallel Bench-top Bioreactor Safety 2.2.1 Unauthorised Persons WARNING! Danger for unauthorised persons! Unauthorised persons are those who do not fulfil the criteria described here and so may not appreci- ate any of the dangers related to operation.

  • Page 16: Conventional Use Of The Equipment

    Multifors 2 - Parallel Bench-top Bioreactor Safety 2.3 Conventional Use of the Equipment The equipment is designed and constructed only for conventional uses as described here. The equipment is conventionally used only as a bioreactor for the culture of: Microorganisms (equipment version for bacterial cultures) ...

  • Page 17: Personal Protective Equipment

    Multifors 2 - Parallel Bench-top Bioreactor Safety 2.4 Personal Protective Equipment It is essential to wear personal protective equipment to minimise health hazards Always wear the personal protective equipment which the par-  ticular activity requires. Always follow instructions available in the workplace, regard- ...
  • Page 18 Multifors 2 - Parallel Bench-top Bioreactor Safety Safety goggles To protect the eyes against liquid splashes Protective gloves (chemical resistant) To protect hands from aggressive substances. Check gloves are impermeable before use. Clean gloves before taking them off and store in a well-ventilated location after use...

  • Page 19: Particular Hazards

    Multifors 2 - Parallel Bench-top Bioreactor Safety 2.5 Particular Hazards The following section contains additional risks which were identi- fied on the basis of a risk assessment. Observe all safety instructions and warning notices in this and the following sections, in order to reduce health hazards and to avoid dangerous situations.
  • Page 20 Multifors 2 - Parallel Bench-top Bioreactor Safety Biohazard WARNING! Biohazard! Noncompliance with biological safety regulations increases the health risk of the operator and the risk of lower equipment functionality. Therefore: – Strictly follow all biological safety regulations – Decontaminate and sterilise every part including...
  • Page 21 Multifors 2 - Parallel Bench-top Bioreactor Safety Dangerous gases WARNING! Risk of explosion, risk of asphyxiation and high risk of danger to the health due to dangerous gases! Non-compliance with safety regulations regarding the use of dangerous gases such as e.g. O...

  • Page 22: Environmental Protection

    Multifors 2 - Parallel Bench-top Bioreactor Safety 2.6 Environmental Protection CAUTION! Danger for the environment caused by mishan- dling! Mishandling environmentally hazardous sub- stances, especially incorrect disposing, may cause severe damage to the environment. Therefore: – Always observe the points stated below.
  • Page 23 Multifors 2 - Parallel Bench-top Bioreactor Safety Biohazards Warns against infectious biochemical substances (e.g. liquids which contain bacteria or viruses) present in the working area. Some procedures must be executed on objects or in rooms which contain bacteria, yeasts or other parasites. These substances may become perilous not only to medical and laboratory personnel, but also to cleaners when handled incorrectly.

  • Page 24: Technical Data

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3 Technical Data 3.1 Equipment Dimensions Dimensions in mm Page 24 of 211 28 March 2018...

  • Page 25: Dimensions Culture Vessels And Vessel Holder

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.2 Dimensions Culture Vessels and Vessel Holder Side View Depending upon height of the culture vessels, the height of the frame which houses them is adjustable. Dimensions Vessels, Total volume 0,4 l Bact./Cell 0,70 l Cell 0,75 l Bact.
  • Page 26 Multifors 2 - Parallel Bench-top Bioreactor Technical Data Top View Dimensions in mm, apply to all vessel sizes. Page 26 of 211 28 March 2018...

  • Page 27: Weights

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.3 Weights Description Value Unit Touch screen operating panel 5 kg Basic unit with 2 culture vessels approx. 40 kg (equipped with standard compo- nents) and operating panel 28 March 2018 Page 27 of 211...

  • Page 28: Service Requirements

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.4 Service Requirements 3.4.1 Electrical Description Type 230 V Type 115 V Unit Voltage Frequency range 50/60 Current 3.4.2 Water IN Description Value Unit Connection pressure 2 ± 1 bar Temperature min.

  • Page 29: Exit Gas

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.4.5 Exit Gas Description Value Unit Connection pressure no back pressure Connection: hose nozzle, outer 5 mm diameter 28 March 2018 Page 29 of 211...

  • Page 30: Specifications

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.5 Specifications 3.5.1 Operating Unit Description Value Colour touch screen Operating system Windows XP Embedded Software INFORS HT parallel bioreactor software for up to 6 individual bioreactors OPC Server OPC XML DA 1.0...
  • Page 31 Multifors 2 - Parallel Bench-top Bioreactor Technical Data Vessel top plate ports bacterial system Port Number / vessel size Thread 400 mL TV 750 mL TV 1400 mL TV 7 mm Without 10 mm Without 12 mm Pg13.5 Vessel top plate ports cell culture system...

  • Page 32: Stirrer Drive System Bacterial Version

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.5.3 Stirrer Drive System bacterial Version Description Value Drive Magnetic Motor Type: DC, external rotor motor Nominal power: 56 W Nominal torque: 90 mNm Range of rotation Vessel 400 mL and 750 mL TV:...

  • Page 33: Stirrer Drive System Cell Culture Version

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data Impellers Number/Type and Material Vessel 2 blade impellers, 6 blades 400 mL 316L, electro polished Ra = 0.8 µm 2 Rushton impellers, 6 blades 750 mL and 1400 mL Peek Vessel 400 mL 24 mm 6.5 mm...
  • Page 34 Multifors 2 - Parallel Bench-top Bioreactor Technical Data Impellers Type und Material Number Vessel Pitched blade impeller, 3 blades 2 pieces 400 mL 316L, electro polished, Ra = 0.8 µm 1 piece 700 mL Angle of 45° producing an upward...

  • Page 35: Temperature

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.5.5 Temperature Description Value Sensor PT-100 1/3 DIN-B Heating Electric, thermal block 315 W With tap water or chiller via thermal Cooling  block (default) With cold finger in 12 mm port (op- ...

  • Page 36: Gassing Bacterial System

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.5.6 Gassing Bacterial System The entire gas entry takes place via the sparger. The specific gassing rate, calculated for the max. working volume is for each vessel size and each variant 2 min...

  • Page 37: Gassing Cell Culture System

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.5.7 Gassing Cell Culture System Basically, the entire gas entry takes place via the sparger. Additionally, head space gassing with air is possible, too. This pre- vents from accumulation of CO in the head room and reduces foam.

  • Page 38: Antifoam

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data Measuring ranges of mass flow controllers and rotamters Max. working Measuring range MFC Measuring range Rota- Measuring range Rotameter volume meter mL min mL min vessel L min (Variants Standard and (sparger, pH control)
  • Page 39 Multifors 2 - Parallel Bench-top Bioreactor Technical Data NOTICE! HAMILTON pH sensors are preconfigured before dispatch by the equipment manufacturer IN- FORS HT. Replacement sensors of this type must be reconfigured before use! NOTIC! Details on technical data, use and maintenance of the pH sensors can be found in the separate docu- mentation of the sensor manufacturer.

  • Page 40: Pumps

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data NOTICE! Details about technical data, use and maintenance of the sensors are in the separate documentation from the sensor manufacturers. 3.5.11 Pumps Description Value Type Peristaltic Standard Digital Acid (3 pieces) Base...

  • Page 41: Sterile Filters

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.5.12 Sterile Filters Description Value All filters Autoclavable Disposable Inlet air filter Retention rate: 0.2 µm (red label) Diameter: 37 mm Inlet air filter Retention rate: 0.2 µm (no label) Diameter: 25 mm Exit gas filter Retention rate: 0.3 µm dry, 1.0 µm wet...

  • Page 42: Operating Conditions

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.6 Operating Conditions Description Value Unit Range of temperature 5 to 40 °C Relative humidity, non-condensing 20 to 90 % 3.7 Emissions Description Value Unit Noise emission <70 dB(A) Page 42 of 211...

  • Page 43: Identification Plate

    Multifors 2 - Parallel Bench-top Bioreactor Technical Data 3.8 Identification Plate The identification plate is located on the bottom left at the back of the basic unit. It contains the following information: Designation: Device designation (category)  Type: Model/Type (Name) ...

  • Page 44: Setup And Function

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4 Setup and Function 4.1 Basic Unit Equipment frame Thermal block Measuring amplifier Vessel holder and support frame Hand grip basic unit (1 x left, 1 x right) Main switch Pumps Two vessels (= bioreactors) of the same size can be connected to a basic unit.
  • Page 45 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function The entire instrumentation and control equipment is integrated into the housing of the basic unit. At the top left and top right of the basic unit there are measuring amplifiers with an integrated temperature sensor (Pt100) and con- necting cables for the pH, pO and antifoam sensors.

  • Page 46: Main Switch

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.1.1 Main Switch The main switch is a green rocker-type. It is located at the top left on the front of the basic unit and is marked "POWER". The main switch is illuminated when the equipment is switched on.

  • Page 47: Water Connections

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.1.3 Water Connections The connections for water feed and return of the basic unit are lo- cated at the centre bottom on the back of the basic unit. They are labelled as follows: "Water out": water outlet...
  • Page 48 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Water inlet of exit gas cooler, respectively water outlet of  basic unit Water outlet of exit gas cooler, respectively water inlet of  basic unit The control valves for the water flow are labelled with an icon The valves are preset in the factory.

  • Page 49: Connection Of The Sparger

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.1.6 Connection of the Sparger The hose nozzles to connect the sparger to the gassing are lo- cated beneath the rotameter mounted on the unit's frame (number depends on equipment configuration).

  • Page 50: Pumps

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.1.7 Pumps Reagents and feed solution are added via four peristaltic pumps. Each culture vessel has four pumps installed in the standard con- figuration. The pumps are driven by stepper motors.
  • Page 51 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Feed solution can also be added via an additional analogue pump. This is mounted on the side of the basic unit. Several free-standing pumps can also be used as external units.

  • Page 52: Operating Panel

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.2 Operating Panel The operating panel has a 12"TFT colour-touch screen. The oper- ating panel is dust-proof and protected against strong water jets, this corresponds to protection class IP66. Up to six bioreactors (= culture vessels) can be individually or in parallel controlled by one operating panel.

  • Page 53: Operating Panel Connections

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function NOTICE! The ON/OFF key is locked when a bioreactor (fer- mentation) is running. A dialogue box with the ap- propriate message appears when pressing the ON/OFF key by mistake during fermentation. When...

  • Page 54: Vessels, Vessel Holder & Transport Frame

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.3 Vessels, Vessel holder & Transport Frame The vessels are made of single-walled borosilicate glass. All ves- sels are delivered with a pre-assembled top plate in a stainless steel vessel holder and transport frame.

  • Page 55: Vessel For Bacterial Cultures

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Two individual vessel holder and transport frames that can be joined together are available as an optional extra. 4.3.1 Vessel for bacterial Cultures Flat-bottom vessels for bacterial cultures are available in three...
  • Page 56 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function The flat-bottom vessels are available with a GL1/4 port at the bot- tom of the vessel as an optional extra. Page 56 of 211 28 March 2018...

  • Page 57: Vessel For Cell Cultures

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.3.2 Vessel for Cell Cultures Round-bottom vessels for cell cultures are available in two sizes: 700 mL total volume / nominal diameter 70  1000 mL total volume / nominal diameter 90 ...

  • Page 58: Baffle Cage

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.3.3 Baffle Cage 1400 mL vessels for bacterial cultures have a baffle cage as a standard fitting. The cage is made in two pieces that are held together with two cheese-head screws and which also secure it to the bearing car- rier.
  • Page 59 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function A baffle cage is also available for 750 mL culture vessels as an op- tion. It is made in one piece and is permanently welded onto the bearing carrier. 28 March 2018...

  • Page 60: Vessel Top Plate Ports And Configuration

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.3.4 Vessel Top Plate Ports and Configuration The vessel top plate has different ports of different sizes to mount the different components such as sparger, blanking plugs, sensors etc. The vessel top plate is equipped with: A clamping ring with quick-release fastener to secure the top ...
  • Page 61 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Blanking plugs with O-ring for the 10 mm ports. Cheese-  head screws are used to secure the blanking plugs 4 addition port adapters with O-rings for reagents  The addition port adapters have an oblique tip. The long sides of the needle tips must all face outwards;...
  • Page 62 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 1 blind tube for the temperature sensor (Pt100) and cheese-  head screw to secure it Holder for reagent bottle hoses  The hose holder is also used to secure the addition port adapters.
  • Page 63 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Magnetic stirrer for bacteria or cell cultures. For details on stir-  rers, see chapter "Drive and Stirrer". 1400 mL vessels for bacterial cultures are all equipped with a baf- fle cage. For details see the chapter “Baffle Cage”.

  • Page 64: Top Plate With Nominal Width 55/70

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.3.4.1 Top Plate with nominal Width 55/70 Ø 12 mm / Pg13.5: Exit gas cooler Ø 7 mm: Addition port adapter (4 pieces Ø 12 mm / Pg 13.5: Sparger Ø 12 mm / Pg13.5: pO sensor Ø...

  • Page 65: Vessel Top Plate With Nominal Width 70

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.3.4.2 Vessel Top Plate with nominal Width 70 Ø 10 mm: Sparger Ground for antifoam sensor Ø 12 / Pg13.5 mm: Exit gas cooler Ø 10 mm: Antifoam sensor Ø 7 mm: Addition port adapter (4 pieces) 12 / Pg 13.5 mm: pH sensor...

  • Page 66: Vessel Top Plate With Nominal Width 90

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.3.4.3 Vessel Top Plate with nominal Width 90 Ø 12 / Pg 13.5 mm: pO sensor Ø 12 / Pg 13.5 mm: pH sensor Ø 10 mm: Sparger Ground for antifoam sensor Ø...

  • Page 67: Temperature Control System

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.4 Temperature Control System Thermal block Clamp (snap closure) The temperature is controlled using a dry system comprising two thermal blocks made of aluminium. Cooling is carried out via a heat-exchange system in the thermal block.

  • Page 68: Drive And Stirrer

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.5 Drive and Stirrer Stirring is carried out from below using magnetic drives. Stepper motors and speed monitoring are used to ensure compliance with the specified stirrer speeds. For speed ranges, see the chapter "Technical Data".

  • Page 69: Drive Shaft For Cell Cultures

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Centring plate of drive shaft Stirrers for bacterial cultures in 1400 mL vessels are equipped with four magnets as well as a centring plate. 4.5.2 Drive Shaft for Cell Cultures Magnet holder...

  • Page 70: Impellers

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.5.3 Impellers The standard stirrer for bacterial cultures has 2 Rushton impellers with 6 blades. The impellers are made of PEEK. Exception for 400 mL vessels: two blade impellers with 6 blades made of 316L are used here.

  • Page 71: Gassing System

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.6 Gassing System Rotameter Pressure reducing valve Manometer The basic unit is equipped with the appropriate gassing units (rota- meter, solenoid valves, mass flow controllers and configured ac- cording to the chosen gassing strategy.

  • Page 72: Gassing Strategy

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function As an option for cell cultures, carbon dioxide (CO ) can be used instead of liquid acid for pH control. 4.6.1 Gassing Strategy The following variants are available: Basic (bacterial system only) Manual flow control via rotameter ...
  • Page 73 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 1400 mL vessels for bacterial cultures  700 mL vessels for cell cultures  1000 mL vessels for cell cultures  Sparger with aeration ring Sparger Aeration ring Centring plate The sparger is connected to an aeration ring. It has a central cen- tring plate in which the location bearing of the drive shaft is placed.
  • Page 74 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function The illustration on the left shows the lower section of a sparger with the aeration ring as well as the drive shaft with location bearing and magnets. Details of the drive shaft are given in chapter "Drive and stirrer".
  • Page 75 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function The illustration on the left shows the lower section of a ring sparger along with the drive shaft with magnets and the impeller used in a vessel for cell cultures. Details of the drive shaft and stirrer are given in chapter "Drive and stirrer".

  • Page 76: Exit Gas And Exit Gas Cooler

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.6.3 Exit Gas and Exit Gas Cooler The exit gas leaves the culture vessel via the thermodynamic exit gas cooler in which the entrained moisture condenses and flows back into the culture vessel.

  • Page 77: Accessory Kit

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.7 Accessory Kit Standard packages for bacterial or cell cultures include an acces- sory kit with the following components: Exit gas cooler (see chapter "Gassing and Exit gas ")  Antifoam sensor, sensor and dosing needle combined ...
  • Page 78 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Type with clamping adapter for 10 mm ports Clamping adapter Dosing needle Round sensor/needle tip The antifoam sensor can be adjusted in its height after loosening the screw on the clamping adapter.
  • Page 79 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Type with clamping adapter for 12 mm ports Hollow screw Housing of clamping screw with thread Dosing needle Sharp sensor/needle tip The antifoam sensor can be adjusted in its height after loosening the hollow screw.

  • Page 80: Reagent Bottles

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function as isolation and must always be intact. A reserve is included in the set. NOTICE! To mount the transparent sheathing, carefully pull it over the dosing needle and shrink coat it by warm- ing it up using e.g.

  • Page 81: Sampling System

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function 4.7.3 Sampling System Basically, different systems and also individual components are available for sampling. As a standard, the aseptic sampling system Super Safe Sampler is delivered with the ready assembled equipment packages for bacte- rial and cell cultures.
  • Page 82 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Elements of the valve assembly Air filter Check valve Luer activated sample valve T-piece Silicone hose The valve assembly consists of a T-piece, two check valves, a Luer-activated automatic sample valve, an air filter, a length of...
  • Page 83 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function NOTICE! Disinfection of the automatic sample valve by wip- ing with a commercial disinfectant can be accom- plished before fitting the syringe, in order to guar- antee the sterility of the sample. This measure is not necessary to ensure the sterility of the bulk cul- ture and can be omitted.
  • Page 84 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Practical tips for the use of the Super Safe Sampler Sterility of the culture vessel is ensured at all times without the pos- sible measures mentioned below. The use of a sterile syringe and sterile caps is only necessary if the sample has to be processed under sterile conditions.
  • Page 85 Multifors 2 - Parallel Bench-top Bioreactor Setup and Function Before fitting the syringe, carry out a wiping disinfection of the sam- ple valve. Spray a commercially available disinfectant onto the valve, leave to soak, and then wipe off with a sterile cloth.

  • Page 86: Starter-Kit

    Multifors 2 - Parallel Bench-top Bioreactor Setup and Function For this proceed as follows: Procedure Remove Adapter 1, check valve, Adapter 2 and sample valve by one left turn of each item. Screw the check valve (without adapter!) with opposing orien- tation directly onto the T-piece.

  • Page 87: Field Of Application

    Precisely defined fermentation conditions enable the execution of parallel experiments. 4.9 Types The equipment is available in four types: Multifors 2 for bacterial, yeast and fungi cultures:  2 x 400 mL total volume, flat bottom vessel  2 x 750 mL total volume, flat bottom vessel ...

  • Page 88: Transport, Packaging And Storage

    Multifors 2 - Parallel Bench-top Bioreactor Transport, Packaging and Storage 5 Transport, Packaging and Storage 5.1 Safety Instructions for Transport Personnel Transport of the equipment may only be carried out by qualified personnel. Inappropriate transportation CAUTION! Damage due to inappropriate transportation! Inappropriate transportation may cause considera- ble loss of property.

  • Page 89: Packaging

    Multifors 2 - Parallel Bench-top Bioreactor Transport, Packaging and Storage 5.3 Packaging The packaging The packaging is composed exclusively of environmentally-friendly materials. The equipment and accessories are packed in a wooden crate. Ac- cessories are additionally packed in cardboard boxes.

  • Page 90: Transport

    Multifors 2 - Parallel Bench-top Bioreactor Transport, Packaging and Storage 5.4 Transport NOTICE! The following descriptions refer to the transport of new, unused equipment. WARNING! Risk of injury due to inappropriate transporta- tion. Inappropriate transportation may lead to severe in- juries and loss of property due to the equipment’s...

  • Page 91: Storage

    Multifors 2 - Parallel Bench-top Bioreactor Transport, Packaging and Storage 5.5 Storage 5.5.1 Storage of the Equipment in General The equipment may only be stored under the following conditions: Do not store outdoors.  Do not expose to aggressive media.

  • Page 92: Storage Of The Reagent Bottles And Hoses

    Multifors 2 - Parallel Bench-top Bioreactor Transport, Packaging and Storage 5.5.2 Storage of the Reagent Bottles and Hoses WARNING! Risk of injury due to inappropriate storage of reagent bottles and hoses! Residues of acid or base may damage the reagent hoses or elements of the bottle caps when storing unwashed reagent bottles and hoses.

  • Page 93: Installation And Initial Operation

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6 Installation and Initial Operation Faulty installation or incorrect WARNING! initial operation Danger due to faulty installation or incorrect in- itial operation! Installation and initial operation requires qualified and experienced personnel. Faulty installation may lead to perilous situations or severe loss of prop- erty.

  • Page 94: Location Requirements For Installation

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation Inappropriate installation and initial WARNING! operation Risk of injury due to inappropriate installation and initial operation! Inappropriate installation and initial operation may lead to severe damage to persons or loss of prop- erty.

  • Page 95: Minimum Spacing

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.2.2 Minimum Spacing To operate and maintain the equipment it must be installed with a minimum spacing of 100 mm from walls, ceilings or other equip- ment. 6.2.3 Supply Lines 6.2.3.1 Air/Gas...

  • Page 96: Exit Gas

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation CAUTION! Danger of loss of property due to use of oily or wet compressed air! Use of oily or wet compressed air may lead to dam- ages to the mass flow controller (option)! Therefore: –...

  • Page 97: Water Inlet

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.2.3.3 Water Inlet There must be a water supply of adequate quality at all times. Cooling water conditions Take the following into consideration in the choice of cooling water: Use only water with no more than a slight hardness for cooling pur- poses.

  • Page 98: Electric Power Supply

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.2.3.5 Electric Power Supply Personnel Connection to be made by a qualified electrician only  Conditions The electric power supply must meet the following conditions: Single phase power supply ...

  • Page 99: Mounting The Operating Panel

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.3 Mounting the Operating Panel The operating panel is fastened with four cylindrical hexagon- socket screws onto the mounting plate on the flanged clamps on the equipment frame. Mounting plate...

  • Page 100: Connecting The Operating Panel To The Basic Unit

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.4 Connecting the Operating panel to the Basic Unit The power supply cable is already connected to the controller in- side the basic unit. During initial operation, the power supply cable only needs to be plugged into the DC socket on the operating panel.

  • Page 101: Connecting The Master Unit With The Satellite Unit(S)

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.5 Connecting the Master Unit with the Satellite Unit(s) Up to 6 bioreactors (= 3 basic units) can be operated and con- trolled in parallel or individually using one operating panel.

  • Page 102: Mounting The Water Connection Set For 1 Or 2 Satellite Units

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.5.1 Mounting the Water Connection Set for 1 or 2 Satellite Units The scope of supply includes a water connection set so that more than 2 bioreactors (= 1 basic unit) can be operated at the same time.
  • Page 103 Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation Connect them as follows: Procedure Insert the push-in hose nozzles into the bulkhead connectors on the plate attached to the master unit. Connect the pressure hoses together using the push-in T-con- nectors .

  • Page 104: Connecting The Controller Of The Master Unit With The Controllers Of The Satellite Unit(S)

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.5.2 Connecting the Controller of the Master Unit with the Controllers of the Satel- lite Unit(s) DANGER! Life-threatening hazard due to electrical cur- rent! Contact with live parts can cause fatal injuries! Therefore: –...
  • Page 105 Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation master unit and the satellite unit(s) can also be identified by the numbering on their identification plate: Serial number and -001 = master unit  Serial number and -002 = 1st satellite unit ...

  • Page 106: Test Run

    Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation 6.6 Test Run Initial operation includes a short test run. This involves re-checking the basic functions of the equipment after installation before users carry out the first fermentation/cultivation. The basic functions are: Temperature control (cooling / heating) ...
  • Page 107 Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation NOTICE! The stirrer cannot start if it is not correctly seated. Drive shafts for cell cultures are not anchored in a bearing. and secure with Carefully fit the top plate on the vessel the clamping ring.
  • Page 108 Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation Check the cooling system: CAUTION! Risk of sustaining burns and loss of property if the thermal block overheats. The thermo block will overheat if the temperature sensor is not fitted to the vessel and/or there is no liquid in the vessel.
  • Page 109 Multifors 2 - Parallel Bench-top Bioreactor Installation and Initial Operation If the heating system is also working, the test is complete. 17. Stop the bioreactor. 18. Shut down the system. 19. Switch off the equipment at the main switch. 20. Shut off the supply lines.

  • Page 110: Preparations For Use

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7 Preparations for Use 7.1 Safety Personnel All preparatory work for use may only be carried out by qualified personnel in biology, biotechnology or chemical engineering Inappropriate preparatory work WARNING! Risk of injury due to inappropriate preparatory...
  • Page 111 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Electric current DANGER! Danger of fatal electric shock! There may be fatal danger by touching compo- nents connected to a mains power supply (single or 3-phase). Damage to insulation or components...

  • Page 112: Preparations Before Sterilisation

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2 Preparations before Sterilisation 7.2.1 Preparing the Vessel WARNING! Risk of injury and loss of property due to dam- aged vessel! A damaged culture vessel (e.g. cracks, scratches, etc.) can burst when under pressure. This can lead...
  • Page 113 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Hold the vessel with both hands on the top plate. Lift the vessel vertically out of the thermal block and put aside on a stable, clean workspace. Removing the vessel top plate...
  • Page 114 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Unhook the threaded bolt. CAUTION! Risk of contamination and danger of loss of property! If the preset length of 21 mm of the spring on the threaded bolt of the closed clamping ring is...
  • Page 115 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Carefully take off the vessel top plate. Check on damages and replace if necessary. Check the glass vessel on damage (cracks, scratches etc.), replace as necessary. Checking gaskets (O-rings) O-rings are used to seal all openings on the vessel and top plate.
  • Page 116 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Check that the O-rings of all component parts and those in the 12 mm ports are correctly seated and undamaged. If neces- sary, reposition or replace. NOTICE! Wet the O-rings with 70% alcohol or a little water to facilitate removing and replacing them or compo- nent parts with fitted O-rings.
  • Page 117 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use A misplaced stirrer cannot start running. Correcting the position of a misplaced stirrer shaft after sterilisation will break sterility of the vessel. NOTICE! Stirrer shafts for cell cultures do not have a location bearing.
  • Page 118 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Adjusting the position of the impellers If necessary, adjust the position of the impellers. For this proceed as follows: Procedure Slightly loosen both grub screws using an Allen key – do not fully unscrew.
  • Page 119 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Fit the clamping ring around the vessel top plate and turn the threaded bolt so that it can be hooked in again. 28 March 2018 Page 119 of 211...
  • Page 120 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Hook in the threaded bolt. Close the clamping ring. Securing the vessel on the basic unit To secure the vessel(s) in the vessel holder and transport frame on the basic unit, proceed as follows: Procedure Insert the vessel in the vessel holder and transport frame.

  • Page 121: Preparing The Sensors

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.2 Preparing the Sensors NOTICE! Initial operation, maintenance and use of the sen- sors are described in detail in the sensor manufac- turer’s documentations. These instructions are strictly to be followed! 7.2.2.1 Calibrating the pH Sensor...
  • Page 122 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use The pH sensor(s) is/are calibrated  Mount the sensors so that they cannot come in contact with  other built-in components or with the glass vessel. Mount the pO sensor so that the bottom cannot collect bub- ...
  • Page 123 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Screw the sensor into the support guide. Insert the sensor into the threaded collar with the thread point- ing in the downward direction. Fit the fork of the rod into the groove of the threaded collar.
  • Page 124 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 10. Screw in an tighten the sensor using the threaded collar. 11. Completely tighten the grub screw in the support guide with the key. : pH! FINALLY Cover connectors of the sensors from the manufacturer METTLER with a little aluminium foil.

  • Page 125: Mounting The Antifoam Sensor

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.2.3 Mounting the Antifoam Sensor Respect the following points, when mounting the antifoam sensor: The sensor head must not touch the clamping adapter, re-  spectively the hollow screw on the clamping adaptor, other- wise a continuous short-circuit is generated, indicating “Foam”.
  • Page 126 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Antifoam sensor with clamping adapter 10 mm ports Loosen the screw next to the blanking plug in the 10 mm port with a screw driver (no. 3) and remove it. Pull out and remove the blanking plug from the port.
  • Page 127 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Antifoam sensor with clamping adapter 12 mm ports Carefully and manually screw the antifoam sensor on the thread of the housing of the clamping screw into the port and tighten it.

  • Page 128: Preparing The Reagent Bottles, Hoses And Pumps

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.3 Preparing the Reagent Bottles, Hoses and Pumps WARNING! Risk of injury due to bursting of corrective rea- gent hoses! Reagent hoses can burst or come off the addition port adapters during sterilisation in the autoclave due to overpressure or damaged hoses.

  • Page 129: Connecting The Reagent Bottles To The Pumps And Vessels

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use A detailed description how to calibrate a pump can be found in the separate touch screen software manual from the equipment manu- facturer. 7.2.5 Connecting the Reagent Bottles to the Pumps and Vessels...
  • Page 130 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Connection of reagent bottle to pumps: Fit both silicone hoses of the reagent bottle onto the hose con- nector of the left pump hose (= intake side, see direction of ro- tation of the pump in the picture on the left) on both pump heads, e.g.
  • Page 131 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 11. Fit the silicone hoses onto the addition port adapters on the vessel top plate. 12. Fit the silicone hoses of the antifoam pumps onto the mounted antifoam sensors in the vessels.

  • Page 132: Preparing The Sampling System Super Safe Sampler

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.6 Preparing the Sampling System Super Safe Sampler WARNING! Danger of health risk when working with patho- genic organisms! Mechanical failure of the automatic valves may cause leakage of culture from the sampling system.
  • Page 133 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use CAUTION! Danger of loss of property due to using force! If the screw connections of the sampling system are tightened with force, this can damage of the component parts. Therefore: – Gently tighten all screw connections with two fingers.
  • Page 134 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Fit the hose of the valve group to the dip tube. The hose can be secured with a cable tie for additional secu- rity. NOTICE! Hoses filled with water during sterilisation in the au- toclave makes for better heat transfer and thereby reduces the contamination risk.
  • Page 135 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Tightening the screw connection check valve/filter: Gently turn the air filter clock-wise. Loosely cover the valve assembly with aluminium foil. The cover protects the air filter against condensation and keeps the surface of the automatic valve sterile after removal from the autoclave.

  • Page 136: Mounting The Exit Gas Cooler

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.7 Mounting the Exit Gas Cooler Respect the following conditions for sterilisation: The exit gas filter must not come off during sterilisation  The exit gas line must be open and may not be clamped off.

  • Page 137: Preparing The Port In The Vessel Top Plate For Inoculation

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.8 Preparing the Port in the Vessel Top Plate for Inoculation The vessel top plate port must be prepared for later inoculation with the syringe. Proceed as follows: Procedure Loosen and remove the blanking plug from the 12 mm port us- ing the hexagon socket spanner provided with the equipment.
  • Page 138 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Fit the blanking plug into the septum collar. Tighten the blanking plug using the hexagon socket spanner provided with the equipment. Page 138 of 211 28 March 2018...

  • Page 139: Preparing The Sparger

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.9 Preparing the Sparger CAUTION! Danger of loss of property from a dirty inlet fil- ter due to leaking of culture liquid! Culture liquid can leak out of the vessel via the...
  • Page 140 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Fit the inlet filter (filter with red label) in the air flow direction on the hose. The red label "INLET" must be on the top. Loosely secure the inlet air filter with a cable tie.

  • Page 141: Attaching The Pump Heads To The Vessel Holder And Support Frame

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.10 Attaching the Pump Heads to the Vessel Holder and Support Frame The pump heads must be removed from the basic unit and at- tached to the vessel holder and support frame. Proceed as follows: Procedure Pull off and remove the cover plate from the pump heads.

  • Page 142: Transport To The Autoclave

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.2.11 Transport to the Autoclave The vessels, reagent bottles and pump heads are now one unit in the vessel holder and support frame and can be transported to the autoclave. Proceed as follows:...

  • Page 143: Sterilisation In The Autoclave

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.3 Sterilisation in the Autoclave The vessel must be custom-designed sterilised in an autoclave be- fore starting the fermentation/cultivation process. WARNING! Microbial danger! An incorrect sterilisation procedure may lead to a...
  • Page 144 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Sterilising the vessel with culture medium method WARNNG! Risk of injury due to splashing liquids when taking a vessel out of the autoclave which is too hot! Superheating and subsequent boiling may occur when sterilising liquids.
  • Page 145 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use CAUTION! Risk of damage to the pH sensor! The glass membrane of the pH sensor can be damaged due to drying out. The water vapour gen- erated by sterilisation is sufficient for moistening it.

  • Page 146: Preparations After Sterilisation

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.4 Preparations after Sterilisation 7.4.1 Mounting the Vessel Holder with the Vessels on the Basic Unit CAUTION! Danger of scalding and burns due to contact with hot surfaces! Touching the vessel holder and support frame, the...
  • Page 147 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Hold the vessel holder and support frame on both hand grips and replace it around the thermal blocks from the top. Engage the vessel retaining brackets, i.e. release the clamps of the snap closure between the thermal blocks.
  • Page 148 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Fit the hoses behind the hose retainer. Page 148 of 211 28 March 2018...

  • Page 149: Filling The Reagent Hoses

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.4.2 Filling the reagent Hoses To prepare the reagent hoses for operation, they must be filled with reagent. Proceed as follows: Procedure Switch the equipment on at the main switch. Open the clamps on the corrective reagent tubes.

  • Page 150: Connecting The Sparger To The Gas Supply

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.4.3 Connecting the Sparger to the Gas Supply The sparger must be connected to the gas supply on the basic unit after sterilisation. Proceed as follows: Procedure Cut off piece of thick-walled silicone hose.

  • Page 151: Connecting The Temperature Sensor

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.4.4 Connecting the Temperature Sensor The temperature sensor (Pt100) has no connector. To connect it, proceed as follows: Procedure Insert the temperature sensor as far as it will go into the pocket in the vessel top plate.

  • Page 152: Testing The Antifoam Pump, Connecting The Antifoam Sensor

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.4.5 Testing the Antifoam Pump, connecting the Antifoam Sensor The function of the antifoam pump can be tested before connecting the cable with the sensor. Proceed as follows: Procedure Switch on the equipment at the main switch.
  • Page 153 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Procedure Insert the red banana connector into the hole in the sensor head. Insert the black banana connector into the ground connector in the vessel top plate. 28 March 2018 Page 153 of 211...

  • Page 154: Connecting The Ph And Po2 Sensors

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.4.6 Connecting the pH and pO2 Sensors CAUTION! Risk of damaging the sensor cables! The integrity of the sensor cables can be damaged by buckling or twisting. This may lead to faulty measurements.
  • Page 155 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Example METTLER pO2 sensor and sensor cable with bayonet coupling. To connect the sensor cable, the red mark on the cable connector and on the sensor connector must be aligned. Example HAMILTON ARC sensor with sensor cable to plug on and screw tight.

  • Page 156: Polarising A (Polarographic) Mettler Po

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.4.7 Polarising a (polarographic) METTLER pO Sensor Polarographic pO sensors from the manufacturer METTLER must be polarised at initial operation or after disconnection from the volt- age source. Correct calibration is not possible otherwise. This means that the pO sensor must be polarised before calibration.

  • Page 157: Connecting The Exit Gas Cooler

    Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use 7.4.9 Connecting the Exit Gas Cooler Water evaporates from the culture medium depending on the gas- sing rate and the working temperature. The exit gas cooler reduces this loss of water as much as possible.
  • Page 158 Multifors 2 - Parallel Bench-top Bioreactor Preparations for Use Loosen the adjustable hose clamp so that the exit gas filter will serve as a predetermined breaking point if necessary. Push the coupling nipples of the pressure hoses into the con- nection on the front side of the basic unit.

  • Page 159: Operation

    Multifors 2 - Parallel Bench-top Bioreactor Operation 8 Operation 8.1 Safety Personnel The equipment may only be operated by qualified personnel in bi- ology, biotechnology or chemical engineering. Inapppropriate operation WARNING! Risk of injury due to inappropriate operation! Inappropriate operation may lead to severe physi- cal injury or loss of property.
  • Page 160 Multifors 2 - Parallel Bench-top Bioreactor Operation Escape of noxious gases WARNING! Explosion and/or danger of suffocation due to escape of dangerous gasses! Leaky or incorrectly fastened hoses on the gas supply (particularly regarding the supply of pure gases such as oxygen or nitrogen!), both from the supply to the equipment and subsequently to the vessel, can lead to the escape of dangerous gases.
  • Page 161 Multifors 2 - Parallel Bench-top Bioreactor Operation Risk of chemical burns WARNING! Risk of chemical burns! Leaky or inappropriately tightened reagent hoses may lead to leakage of acids and bases. Contact with acids and bases carries a high risk of chemical burns.

  • Page 162: Starting Fermentation

    Multifors 2 - Parallel Bench-top Bioreactor Operation 8.2 Starting Fermentation WARNING! Risk of injury caused by overpressure in the vessel! A cracking vessel or bursting corrective reagent hoses due to overpressure in the vessel can lead to severe injuries and loss of property! Therefore: –...

  • Page 163: Switching On The Equipment

    Multifors 2 - Parallel Bench-top Bioreactor Operation 8.2.1 Switching on the Equipment To switch on the equipment, proceed as follows: Procedure Switch the equipment on at the main switch. The operating unit switches on automatically and the system boots up.

  • Page 164: Inoculation

    Multifors 2 - Parallel Bench-top Bioreactor Operation 8.2.2 Inoculation There are different possible methods of inoculation: Concentrated in a small volume, with the syringe via the sep-  tum into the vessel. With the inoculation needle via the septum with collar into the ...
  • Page 165 Multifors 2 - Parallel Bench-top Bioreactor Operation For inoculation proceed as follows: Procedure Plug the needle onto the syringe and draw back the syringe plunger to fill it with the necessary volume of inoculum. Unscrew and remove the blanking plug out of the septum col- lar.

  • Page 166: Sampling

    Multifors 2 - Parallel Bench-top Bioreactor Operation 8.2.3 Sampling A sample is taken out of the culture medium to gain data for the offline analysis The method of sampling can vary due to the different user applica- tions. 8.2.4 Sampling with Sampling System Super Safe Sampler...
  • Page 167 Multifors 2 - Parallel Bench-top Bioreactor Operation Proceed as follows: Procedure If present, remove the clamp from the sample hose. Check all screw connections on the valve assembly for tight- ness. Carefully tighten the screw connections with two fingers as necessary.
  • Page 168 Multifors 2 - Parallel Bench-top Bioreactor Operation NOTICE! Only use a dry and clean syringe in order to avoid clogging of the filter. This syringe can be reused as often as desired because the air is led via a sterile filter.
  • Page 169 Multifors 2 - Parallel Bench-top Bioreactor Operation Procedure Completely pull back on the piston of the sample syringe. Ex- cept for a few µl, all remaining liquid is removed. While holding the sample valve with one hand, use the other hand to loosen the syringe by turning it a quarter-turn counter clockwise.

  • Page 170: Ending Fermentation

    Multifors 2 - Parallel Bench-top Bioreactor Operation 8.3 Ending Fermentation CAUTION! Risk of injury due to uncontrolled restart of the stirrer and peristaltic pumps. Stopping fermentation by the main switch is like a power cut. I.e. it leads to resuming the former set- tings when switching on again.

  • Page 171: Harvest

    Multifors 2 - Parallel Bench-top Bioreactor Operation 8.3.1 Harvest CAUTION! Danger for the environment caused by mishan- dling! Mishandling environmentally hazardous sub- stances, especially incorrect disposing, may cause severe damage to the environment. Therefore: – Dispose of or re-use biological dangerous or- ganisms according to the local regulations and current protocols in use in the laboratory.
  • Page 172 Multifors 2 - Parallel Bench-top Bioreactor Operation WARNING! Risk of uncontrolled leakage of microorgan- isms! Build-up of excessive pressures in the vessel can cause hoses to detach or burst. Therefore: – Never allow the pressure to exceed the required overpressure.

  • Page 173: Shutting Down The System, Switching Off The Equipment

    Multifors 2 - Parallel Bench-top Bioreactor Operation 8.3.2 Shutting down the System, switching off the Equipment Once harvesting is complete and the reagent hoses are empty, the system can be shut down and the equipment switched off. Proceed as follows:...

  • Page 174: Maintenance

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 9 Maintenance 9.1 Safety Personnel The here described maintenance work can be carried out by  the user if not indicated otherwise (see “Maintenance plan”). Any work on the electrical system is strictly to be carried out ...
  • Page 175 Multifors 2 - Parallel Bench-top Bioreactor Maintenance Electrical system DANGER! Danger of fatal electric shock! Contact with electrically powered components is perilous. Energised electrical components may per- form uncontrolled movements which can lead to severe injuries. Therefore: – Turn off the electrical supply and check whether it is electrically isolated before starting any work.

  • Page 176: Environmental Protection

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 9.2 Environmental Protection CAUTION! Environmental danger by inappropriate han- dling! Inappropriate handling of environmentally hazard- ous substances, especially where disposal is in- volved, may lead to severe environmental damage. Therefore: – Immediately take appropriate action if environ- mentally hazardous substances are accidentally released into the environment.
  • Page 177 Multifors 2 - Parallel Bench-top Bioreactor Maintenance Interval Maintenance work To carry out by Before every op- Check supply lines and connection hoses for leaks and replace as User eration necessary Check all O-rings and rubber seals and replace if necessary...

  • Page 178: Sterilising The Vessel After Fermentation

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 9.4 Sterilising the Vessel after Fermentation The vessel, the reagent bottles and hoses including the pump heads and all components that have been in contact with the me- dium must be sterilised again in the autoclave after the harvest process and before cleaning.
  • Page 179 Multifors 2 - Parallel Bench-top Bioreactor Maintenance CAUTION! Risk of damage to the pump heads! Sterilising the pump heads at too high tempera- tures may lead to damage. Therefore: – Never sterilise the pump heads at temperatures above 125 °C.

  • Page 180: Multifors 2 Cell: Exchanging The Ball Bearings Of The Stirrer Shaft

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 9.5 Multifors 2 Cell: Exchanging the Ball Bearings of the Stirrer Shaft Because the bearings of the stirrer shaft (cell culture version) are not lubricated, they will wear over time. For this reason, they must be replaced at regular intervals (also see the maintenance plan): After max.

  • Page 181: Dismantling The Used Ball Bearings

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 9.5.1 Dismantling the used Ball Bearings To dismantle the used ball bearings proceed as follows: Procedure Remove the vessel top plate from the vessel and carefully put it top side down. Avoid any stress or knocks on the stirrer shaft! Dismantle the sparger.
  • Page 182 Multifors 2 - Parallel Bench-top Bioreactor Maintenance The O-ring is loose now, and the stirrer shaft will also loosen itself from the bearing holder. Carefully pull the stirrer shaft out of the bearing holder. Page 182 of 211 28 March 2018...
  • Page 183 Multifors 2 - Parallel Bench-top Bioreactor Maintenance The whole stirrer shaft with impeller(s) and the two ball bear- ings comes free now. Loosen – do not remove! – the grub screw on the locking ring with the Allen key. Remove the locking ring and put aside.
  • Page 184 Multifors 2 - Parallel Bench-top Bioreactor Maintenance Remove the O-ring from the stirrer shaft. Pull the upper ball bearing (1) off the distance sleeve (2) on the stirrer shaft. Pull the distance sleeve off the stirrer shaft. Page 184 of 211...

  • Page 185: Mounting The New Ball Bearings

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 10. Pull the lower ball bearing off the stirrer shaft. 9.5.2 Mounting the new Ball Bearings CAUTION! Risk of damaging the ball bearings due to inap- propriate mounting! The ball bearings are damaged and will therefore be unusable, if they are inappropriately mounted or have an incorrect axial loading.
  • Page 186 Multifors 2 - Parallel Bench-top Bioreactor Maintenance View A:  Ball bearing with broad gap (a) between the inner and the outer ring facing upwards View B:  Ball bearing with narrow gap (b) between the inner and the outer ring facing upwards...
  • Page 187 Multifors 2 - Parallel Bench-top Bioreactor Maintenance To mount the new ball bearings, proceed as follows: Procedure Carefully plug the lower ball bearing with the broad gap fac- ing upwards (View A) to the stirrer shaft. NOTICE! The ball bearings only support a one-sided axial load.
  • Page 188 Multifors 2 - Parallel Bench-top Bioreactor Maintenance Carefully insert the upper ball bearing with the narrow gap facing upwards (View B) onto the distance sleeve. NOTICE! The ball bearings only support a one-sided axial load. They absolutely must be mounted with the...
  • Page 189 Multifors 2 - Parallel Bench-top Bioreactor Maintenance Place the locking ring with the small locating ring facing to the bearing.  Push down the locking ring and tighten the grub screw with the Allen key at the same time. 28 March 2018...
  • Page 190 Multifors 2 - Parallel Bench-top Bioreactor Maintenance Carefully insert the stirrer shaft in alignment with the bearing holder into the bearing holder. Never insert the stirrer shaft with force and/or misaligned into the bearing holder – the ball bearings can easily get damaged! Carefully insert the O-ring into the groove on the inside of the bearing holder with.

  • Page 191: Cleaning

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 9.6 Cleaning DANGER! Danger of fatal electric shock! There may be fatal danger by touching electrically powered components. Cleaning when equipment is switched on may have fatal consequences. Therefore: – Disconnect the equipment from the mains sup- ply and check whether it is electrically isolated before cleaning.
  • Page 192 Multifors 2 - Parallel Bench-top Bioreactor Maintenance CAUTION! Environmental hazard due to incorrect han- dling! Risk of extensive environmental damage when handling environmentally hazardous substances in- correctly especially when disposing inappropriately. Therefore: – Respect the local regulations concerning dis- posal of environmentally hazardous substances.

  • Page 193: Vessel

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 9.6.1 Vessel CAUTION! Danger of damaging the stirrer shaft! Components parts of the stirring system may come off and become deformed when dismantling the vessel top plate. Any deformation of the stirrer shaft can lead to damage of the bearing.

  • Page 194: Vessel Top Plate

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance NOTICE! It is recommended to warm the 0.1 N NaOH up to 60 °C and to prolong the duration of stirring for dealing with persistent remains of foam or protein. Stop the bioreactor and closedown the system Switch off the equipment.

  • Page 195: Exit Gas Cooler

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance Procedure Carefully rinse the vessel top plate and its component parts (sparger, stirrer etc.) with distilled water. Use 0.1 N caustic soda solution and afterwards distilled water as necessary. Check the vessel top plate sealing and O-rings on component...

  • Page 196: Sensors

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance A damaged pump head must be replaced inclusive the pump  hose section and vice versa. To clean the reagent bottles and hoses, proceed as follows: Procedure Withdraw the reagent hoses from the bottles and the vessel top plate without dismantling the pump heads.

  • Page 197: Operating Unit

    Multifors 2 - Parallel Bench-top Bioreactor Maintenance 9.6.6 Operating Unit DANGER! Risk of fatal electric shock! There may be fatal danger by touching electrically powered components. Cleaning when the equip- ment is switched on may have fatal consequences. Therefore: – Disconnect all electrical connections to the op- erating unit before cleaning.

  • Page 198: Interferences

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10 Interferences Possible reasons for interference or a problem and how to resolve it is described in the following section. Reduce the service intervals if interferences/problems become in- creasingly common. Contact the manufacturer in case of developing problems which cannot be resolved by following the instructions.

  • Page 199: Tables Of Interferences

    Multifors 2 - Parallel Bench-top Bioreactor Interferences Action in case of interferences The following instructions are to be followed: Immediately switch off the equipment and unplug the mains plug in case of malfunctions which represent an imminent danger to persons or property.

  • Page 200: Interferences Basic Unit And Operating Panel

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10.3 Interferences Basic Unit and Operating Panel Interference Equipment does not work. Power indicator of the main switch is not illuminated; monitor of the operating unit remains dark. Possible cause Remedy Power cable not connected.

  • Page 201: Interferences Drive System

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10.1 Interferences Drive System Interference Stirrer does not start. Possible cause Remedy Parameter Stirrer is not activated. Activate the parameter. Operator Setpoint of parameter Stirrer = 0. Set setpoint > 0. Operator Parameter pO...

  • Page 202: Interferences Temperature Control System

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10.2 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 heating or inadequate heating: thermal block in the basic unit does not get hot even with a high temper-...

  • Page 203: Interferences Ph-System

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10.3 Interferences pH-System Interference No display or incorrect display of pH Possible cause Remedy Sensor cable not connected or not Connect properly if necessary. Operator properly connected. Activate this function in option Setpoint of parameter...
  • Page 204 Multifors 2 - Parallel Bench-top Bioreactor Interferences Interference pH value drifts up and down over time or acid and base are added almost continuously in turn. Possible cause Remedy Incorrect PID setting in pH parame- Check the PID settings and adjust as necessary.

  • Page 205: Interferences Po System

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10.4 Interferences pO System Interferences No display or incorrect display of pO Possible cause Remedy Sensor cable not connected or not Connect properly if necessary. Operator properly connected. pO2 sensor is not calibrated Calibrate the pO sensor.

  • Page 206: Interferences Antifoam Or Level Sensor And Antifoam Pump

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10.5 Interferences Antifoam or Level Sensor and Antifoam Pump Interference Foam/medium is not detected. Possible cause Remedy Check connections and connect properly as neces- Sensor is not properly connected. Operator sary. Interference Foam/medium is always/frequently detected.

  • Page 207: Interferences Feed And Pump

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10.6 Interferences Feed and Pump Interference No feed solution or inadequate feed solution. Possible cause Remedy Parameter Feed is not activated. Activate the parameter. Operator Setpoint of parameter Feed = 0. Set setpoint > 0.

  • Page 208: Returning For Repair

    Multifors 2 - Parallel Bench-top Bioreactor Interferences 10.7 Returning for Repair The operator must return the equipment or the faulty component part(s) to the manufacturer if, after consulting the service depart- ment of the local dealer or the manufacturer, on-site diagnosis and/or repair is not possible.

  • Page 209: Disassembly

    Multifors 2 - Parallel Bench-top Bioreactor Disassembly 11 Disassembly The equipment must be disassembled and disposed of in an envi- ronmentally-friendly manner if it is not in use anymore. 11.1 Safety Personnel Disassembly may only be carried out by qualified personnel ...

  • Page 210: Disassembly

    Multifors 2 - Parallel Bench-top Bioreactor Disassembly Biological safety WARNING! Danger to biological safety! Non-compliance with all biological safety regula- tions signifies an increased health risk to the user and the functionality of the equipment. Therefore: – Strictly follow all biological safety regulations –...

  • Page 211: Disposal

    Multifors 2 - Parallel Bench-top Bioreactor Disassembly 11.3 Disposal Recycle disassembled components, if no agreement is made con- cerning reclaim or disposal. Send metals for scrap  Send plastic components for recycling  Sort and dispose of the remaining components according their ...

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