Operating Manual 2019-04-11 Techfors-S In Situ Sterilisable Pilot Bioreactor / Vessel Volume: 15 L Serial No: S-000129020 Dok-ID: 4820 / KA129689 / V.01.00 / Original Infors AG Headoffice, Schweiz Rittergasse 27 CH-4103 Bottmingen T +41 (0)61 425 77 00 F +41 (0)61 425 77 01 info@infors-ht.com service@infors-ht.com Infors GmbH Dachauer Str. 6 D-85254 Einsbach T +49 (0)8135 8333 F +49 (0)8135 8320 infors.de@infors-ht.com Infors UK Ltd The Courtyard Business Centre Dovers Farm, Lonesome Lane, Reigate Surrey, RH2 7QT, UK T +44 (0)1737 22 31 00 F +44 (0)1737 24 72 13 infors.uk@infors-ht.com Infors Sarl 2, rue du Buisson aux Fraises Bâtiment D13 F-91300 Massy T +33 (0)1 69 30 95 04 F +33 (0)1 69 30 95 05 infors.fr@infors-ht.com www.infors-ht.fr Infors Benelux BV Markweg 9-A, NL-6883 JL Velp (GLD) P.O. Box 125, NL-6880 AC Velp (GLD) T +31 (0)26 369 31 00 F +31 (0)26 369 31 09 infors.bnl@infors-ht.com Infors USA Inc. 9070 Junction Drive, Suite D Annapolis Junction, MD20701 T +1 301 362 3710 / T +1 855 520 7277 (toll-free USA) F +1 301 362 3570 infors.usa@infors-ht.com www.infors-ht.us Infors Canada 8350 rue Bombardier Anjou, Quebec Canada H1J 1A6 T +1 514 352 5095 F +1 514 352 5610 infors.ca@infors-ht.com Infors Bio-Technology (Beijing) Co., Ltd. Room 505C, Building 106 Lize Zhongyuan Wangjing New Industrial Zone Chaoyang District, Beijing 100102 P.R. of China T +86 10 51652068 F +86 10 64390585 info@infors-ht.com.cn Infors South East Asia 16, 1st Floor, Taman City MY-51200 Kuala Lumpur Malaysia T +603 625 771 81 F +603 625 067 48 info@infors-ht.com.my Infors LATAM Ltda. Rua Dr. Alceu de Campos Conjunto 205 CEP: 04544-000 São Paulo – SP Brasil T +55 (11) 95304-0201 F +55 (11) 98585-5334 Infors.br@infors-ht.com Contact details of our local dealers worldwide can be found on our website. www.infors-ht.com Engineering and production in Switzerland Techfors-S – Vessel Volume 15 L – Serial No. 000129020 Table of Contents 11 April 2019 Page 1 of 257 1 General Information...............................................................8 1.1 About this Manual ..........................................................8 1.2 Explanation of Special Notices ......................................9 1.2.1 Warning Notices .............................................9 1.2.2 Other Notices..................................................9 1.3 Equipment Identification ..............................................10 1.3.1 Identification Plate ........................................10 1.3.2 Plant Identification Plate...............................10 1.3.3 Vessel Identification Plate ............................11 1.4 Declaration of Conformity ............................................11 1.5 Customer Service and Services ..................................11 2 Safety and Responsibility ...................................................13 2.1 Intended Use, Incorrect Use and Misuse ....................13 2.2 Qualified Personnel .....................................................14 2.2.1 Provider ........................................................14 2.2.2 User..............................................................14 2.2.3 Operator .......................................................15 2.3 Unauthorised Persons .................................................16 2.4 Responsibility of the Provider ......................................16 2.5 General Hazards..........................................................16 2.5.1 Electrical Current..........................................17 2.5.2 Unauthorised Spare Parts and Accessories 17 2.6 Particular Hazards .......................................................17 2.6.1 Hot Surfaces.................................................18 2.6.2 Dangerous Gases ........................................18 2.6.3 Flammable or Explosive Substances ...........18 2.6.4 Corrosive or Toxic Substances ....................18 2.6.5 Bioactive Substances or Pathogenic Organisms ....................................................19 2.6.6 Overpressure or Vacuum .............................19 2.6.7 Steam ...........................................................19 2.7 Warning Symbols on the Equipment ...........................19 2.8 Declaration of Decontamination...................................20 3 Setup and Function..............................................................21 3.1 Equipment - Overview .................................................21 3.2 Instrumentation Cabinet...............................................23 3.2.1 Main Switch ..................................................23 3.2.2 Pumps ..........................................................24 3.3 Touch Screen Operating Panel ...................................24 3.3.1 Monitor Keys.................................................25 Techfors-S – Vessel Volume 15 L – Serial No. 000129020 Table of Contents Page 2 of 257 11 April 2019 3.3.2 Operating Panel Connections.......................26 3.4 Vessel – Overview .......................................................27 3.4.1 Vessel Top Plate...........................................28 3.4.2 Vessel Flange...............................................31 3.4.3 Vessel Jacket................................................33 3.4.4 Standard INGOLD Ports...............................33 3.4.5 Vessel Accessories ......................................34 3.5 Harvest/Sample Valve (Bottom Valve).........................35 3.6 Stirrer ...........................................................................38 3.7 Temperature Control & Sterilisation.............................41 3.8 Gassing ........................................................................43 3.9 Exit Gas........................................................................45 3.10 pH Control ....................................................................46 3.11 pO2 Control...................................................................47 3.12 Antifoam Control ..........................................................48 3.13 Transport Lock .............................................................50 3.14 Safety Devices .............................................................51 4 Accessories, Consumables and Hoses .............................53 4.1 Connection Kit..............................................................54 4.2 Manometer (Pressure Indication Vessel).....................54 4.3 Reagent Bottle .............................................................55 4.4 Push Valve...................................................................56 4.5 Sensor Insertion Housing.............................................57 4.6 Inoculation Needles......................................................57 4.6.1 Inoculation Needle, Single............................58 4.6.2 Inoculation Needle, Double ..........................59 5 Transport and Storage.........................................................60 5.1 Transport......................................................................60 5.2 Storage.........................................................................60 6 Installation and Initial Operation ........................................61 6.1 General Location Requirements for Installation...........61 6.2 Minimum Distances......................................................62 6.3 Power Supply...............................................................62 6.4 Cooling Water, Drain & Condensate............................63 6.5 Gas Supply...................................................................64 6.6 Exit Gas........................................................................64 6.7 Steam Supply...............................................................65 7 Preparations before Fermentation / Cultivation................66 7.1 Securing the Position of the Equipment.......................66 7.2 Uncoupling the Motor...................................................67 Techfors-S – Vessel Volume 15 L – Serial No. 000129020 Table of Contents 11 April 2019 Page 3 of 257 7.3 Removing the Exit Gas Cooler and the Filters ............68 7.4 Removing the Vessel Top Plate ..................................70 7.5 Checking Impellers, Baffles and Top Plate Seal .........72 7.6 Filling the Vessel & Refitting the Vessel Top Plate......73 7.7 Checking Lubrication of the Mechanical Seal..............75 7.8 Coupling the Motor ......................................................76 7.9 Preparing the Sensors .................................................77 7.9.1 Mounting / Connecting the Temperature Sensor ..........................................................77 7.9.2 Calibrating the pH-Sensor............................77 7.9.3 Mounting the pH Sensor and Connecting the Cable ............................................................77 7.9.4 Mounting the pO2 Sensor ............................80 7.9.5 Connecting the Sensor Cable ......................81 7.9.6 Polarising the pO2 Sensor (METTLER)........81 7.9.7 Calibrating the pO2 Sensor..........................82 7.9.8 Preparing the Antifoam Sensor ....................82 7.9.9 Mounting the Antifoam Sensor and Connecting the Cable...................................84 7.10 Mounting the Safety Valve...........................................85 7.11 Mounting the Manometer.............................................87 7.12 Calibrating the Pumps .................................................88 7.13 Preparing Reagent Bottles and Inoculation Needles...88 7.13.1 Preparing and Autoclaving ...........................88 7.13.2 Connecting to the Vessel and the Pump......90 7.14 Preparing Push Valves ................................................94 7.14.1 Autoclaving...................................................94 7.14.2 In Situ Sterilisation........................................95 7.14.3 Fermentation/Cultivation ..............................95 7.15 Equipping the Ports with Septa (Piercing Membranes) and Collars...................................................................95 7.16 Closing Unused Ports ..................................................96 7.17 Mounting the Exit Gas Cooler and the Filters..............98 7.18 Checking the Inlet Air/Gas and Exit Gas Lines and Filters ...........................................................................99 7.19 In situ Sterilisation – General Information..................100 8 Fermentation / Cultivation.................................................102 8.1 Preparing the Medium ...............................................102 8.2 Sampling....................................................................103 8.3 Inoculation..................................................................105 8.3.1 Inoculation with Inoculation Needle............105 8.3.2 Inoculation with Push Valve .......................106 8.4 Harvest.......................................................................107 Techfors-S – Vessel Volume 15 L – Serial No. 000129020 Table of Contents Page 4 of 257 11 April 2019 8.5 Emptying the Vessel ..................................................107 8.6 Sterilisation after Fermentation / Cultivation ..............107 9 Operation Touch Screen Software ...................................109 9.1 Screen, Menu Navigation and Control Elements.......109 9.1.1 Screen Areas..............................................110 9.1.2 Control Elements ........................................111 9.2 Main Menus................................................................116 9.2.1 Main – Overview.........................................116 9.2.2 Batch – Start Menu.....................................119 9.2.3 Controller – Value Display..........................120 9.2.4 Cascade......................................................122 9.2.5 Trends – Trend Lines .................................123 9.2.6 System – System Settings..........................125 9.2.6.1 Statistics – Software Communication with Bioreactor Hardware.................................... 126 9.2.6.2 Update – Installing a Software Update........................................ 126 9.2.7 Alarms – Parameter Alarms, User Alarms, System Alarms............................................129 9.2.7.1 Parameter Alarms ...................... 130 9.2.7.2 System-Alarm “Difference in board configuration” ................... 131 9.2.7.3 System Alarm “No water detected in temperature control system”....................................... 131 9.3 Submenus ..................................................................133 9.3.1 Valves – Digital Controller Outputs.............133 9.3.2 Security – User Management.....................135 9.3.2.1 User Levels ................................ 136 9.3.2.2 Access Authorisations of User Groups........................................ 137 9.3.2.3 Login – Logging on to the System ....................................... 141 9.3.2.4 Logout – Logging off the System 142 9.3.2.5 Change Password...................... 143 9.3.2.6 New User – Adding a New User 144 9.3.2.7 Remove User – Deleting a User 146 9.3.2.8 Edit User – Editing User Settings147 9.3.2.9 Set / Delete Default User – Setting or Deleting an automatic User Login.................................. 149 9.3.2.10 Password Security – Configuring Password Rules ......................... 151 9.3.3 Settings – Basic Unit Settings ....................153 Techfors-S – Vessel Volume 15 L – Serial No. 000129020 Table of Contents 11 April 2019 Page 5 of 257 9.3.3.1 IP Settings – Network Settings ...154 9.3.3.2 Change Time – Changing Date and Time .....................................156 9.3.3.3 Backup – Saving Data ................157 9.3.3.4 Restore – Restoring Saved Data or Restoring Factory Settings .....159 9.3.3.5 Export Logs – Exporting Log Files.............................................161 9.3.3.6 Balance Settings .........................162 9.3.4 Wipe Screen – (Temporarily) Locking the Screen ........................................................164 9.3.5 Shutdown – Shutting Down the System.....165 9.4 Recipes ......................................................................166 9.4.1 Save Recipe – Saving a Recipe.................166 9.4.2 Load/Start Recipe – Loading and a Recipe167 9.4.3 Delete Recipe – Deleting a Recipe ............169 9.5 Parameters – Short Description.................................171 9.5.1 Temp ..........................................................171 9.5.2 Stirrer..........................................................171 9.5.3 pH...............................................................171 9.5.4 pO2.............................................................172 9.5.5 Antifoam .....................................................172 9.5.6 Feed ...........................................................172 9.5.7 GasMix .......................................................172 9.5.8 Flow............................................................174 9.6 Parameter Options.....................................................175 9.6.1 Setpoint ......................................................176 9.6.1.1 Setting Setpoint Values, Switching Parameters ON / OFF 178 9.6.1.2 Setting Alarm Values and Critical Values .........................................182 9.6.2 Calibrate .....................................................184 9.6.3 Calibration Menus pH and pO2 Sensors....185 9.6.3.1 Calibrating the pH Sensor – General Information ....................188 9.6.3.2 Calibrating the pH Sensor - Procedure....................................188 9.6.3.3 Recalibrating a METTLER pH Sensor.........................................192 9.6.3.4 Polarising the pO2 Sensor (METTLER).................................192 9.6.3.5 Calibrating the pO2 Sensor – General Information ....................193 9.6.3.6 Calibrating the pO2 Sensor - Procedure....................................193 Techfors-S – Vessel Volume 15 L – Serial No. 000129020 Table of Contents Page 6 of 257 11 April 2019 9.7 PID (Control) ..............................................................197 9.7.1 Explanations of PID Control .......................199 9.7.2 Changing the PID Controller Settings ........200 9.8 Cascade Control ........................................................201 9.8.1 Setting a Cascade ......................................203 9.8.2 Deleting a Cascade ....................................206 9.8.3 Negative Function of a Cascade ................207 9.8.4 Special Configurations................................207 9.9 Pumps........................................................................208 9.9.1 Calibrating a Pump .....................................209 9.9.2 Resetting the Pump Counter to Zero..........211 9.9.3 Automatically Filling/Emptying Pump Hoses ....................................................................212 9.10 SIP – Sterilisation in Place.........................................214 9.10.1 Full Sterilisation – Vessel Sterilisation........214 9.10.1.1 Process Sequences ................... 215 9.10.1.2 Process Configuration................ 216 9.10.1.3 Process Start.............................. 216 9.10.1.4 Process End............................... 218 9.10.1.5 Process Abortion........................ 218 9.10.2 SIP Harvest/Sample Valve – Sterilisation of the Harvest/Sample Valve ..........................219 9.10.2.1 Process Sequences and Process Configuration.............................. 219 9.10.2.2 Process Start and End ............... 219 9.10.2.3 Process Abortion........................ 221 9.11 Starting & Stopping the Bioreactor, Shutting Down the System .......................................................................222 9.11.1 Starting the Bioreactor (Fermentation/Cultivation) ..........................222 9.11.2 Stopping the Bioreactor ..............................225 9.11.3 Shutting Down the System, Switching Off the Equipment...................................................225 10 Cleaning and Maintenance................................................227 10.1 Cleaning Agent and Disinfectant................................227 10.2 Cleaning the Vessel ...................................................227 10.3 Cleaning the Vessel Top Plate...................................228 10.4 Dismantling and Cleaning the Exit Gas Cooler..........229 10.5 Cleaning Reagent Bottles, Hoses and Component Parts ...................................................................................231 10.6 Cleaning the Sensors.................................................232 10.7 Cleaning Surfaces of the Instrumentation Cabinet and Operating Unit ............................................................232 10.8 Maintenance Plan ......................................................233 Techfors-S – Vessel Volume 15 L – Serial No. 000129020 Table of Contents 11 April 2019 Page 7 of 257 10.9 Lubricating the Mechanical Seal................................234 11 Interferences.......................................................................236 11.1 Interferences Basic Operation and Operating Unit....237 11.2 Interferences Drive System .......................................238 11.3 Interferences Temperature Control System ..............239 11.4 Interferences pH System ...........................................240 11.5 Interferences pO2 System (Dissolved Oxygen) ........241 11.6 Interferences Antifoam Control..................................242 11.7 Interferences Feed and Pump ...................................243 11.8 Interferences Gassing System...................................244 11.9 Returning for Repair ..................................................245 12 Disassembly and Disposal................................................246 12.1 Disassembly...............................................................246 12.2 Disposal .....................................................................247 13 Technical Data....................................................................248 13.1 Dimensions, Connections & Connection Values .......248 13.2 Electrical Power Supply.............................................249 13.3 Specifications.............................................................250 13.3.1 Instrumentation Cabinet .............................250 13.3.2 Operating Unit ............................................250 13.3.3 Vessel.........................................................250 13.3.4 Temperature...............................................252 13.3.5 Stirrer..........................................................252 13.3.6 Gassing ......................................................253 13.3.7 Exit Gas......................................................254 13.3.8 pH...............................................................254 13.3.9 pO2.............................................................255 13.3.10 Antifoam .....................................................255 13.3.11 Pumps ........................................................256 13.3.12 Push Valve .................................................256 13.3.13 Pressure Indication Vessel.........................256 13.3.14 Pressure: Safety Valves.............................256 13.4 Operating Conditions .................................................257 13.5 Operating Materials ...................................................257 13.6 Emissions...................................................................257 Techfors-S – Vessel Volume 15 L – Serial No. 129020 General Information Page 8 of 257 11 April 2019 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 technological and scientific developments and the knowledge gained from the manufacturer’s many years of experience in this field. This operating 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. The users must read the operating manual thoroughly and fully understand its contents before beginning any work. Adhering to all the safety and operating instructions in this manual is essential to ensure that work is carried out safely. The scope of delivery may differ from the explanations, descriptions and figures in this operating manual due to special designs, additional options specified on ordering and the latest technical/mechanical modifications. This manual contains illustrations to aid general understanding. These may differ from the actual equipment as supplied. Techfors-S – Vessel Volume 15 L – Serial No. 129020 General Information 11 April 2019 Page 9 of 257 1.2 Explanation of Special Notices 1.2.1 Warning Notices Warning notices in this manual are indicated by a coloured bar and begin with a signal word that signifies the degree of the hazard. DANGER The signal word “DANGER” indicates a dangerous situation that will lead to severe or even fatal injuries if not avoided. 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. 1.2.2 Other Notices ATTENTION The word “ATTENTION” on a blue bar indicates a situation that may result in significant damage to property if not avoided. INFORMATION Texts located below a grey bar bearing the notice “INFORMATION” provide useful tips and recommendations for ensuring efficient, fault-free operation of the equipment. Techfors-S – Vessel Volume 15 L – Serial No. 129020 General Information Page 10 of 257 11 April 2019 1.3 Equipment Identification 1.3.1 Identification Plate The identification plate is designed to allow clear identification of the equipment. It contains the following information:  Manufacturer name  Designation = Category of equipment  Type = Equipment type (name)  S/N = Serial number  Year = Year of manufacture  Mains = Nominal voltage and frequency  Current = Current consumption  Manufacturer address  CE marking 1.3.2 Plant Identification Plate The plant identification plate is located on the central pillar of the basic unit and contains the following information:  Manufacturer name with address  Designation of plant  Equipment type (Name)  Serial number  Year of manufacture  CE marking  Process range (bar) in process area and temperature control system  Temperature range (°C) in process area and temperature control system  Test date Techfors-S – Vessel Volume 15 L – Serial No. 129020 General Information 11 April 2019 Page 11 of 257 One identification plate can be found on the instrumentation cabinet and one on the support of the basic unit. 1.3.3 Vessel Identification Plate The vessel identification plate is welded onto the outside of the vessel. It contains the following data:  Manufacturer of the equipment with address  Manufacturer of the vessel with address  Number of manufacture / Year of manufacture It also contains different values for vessel and vessel jacket:  Max. operating pressure  Max. operating temperature  Total volume (litres)  Material  Pressure test  Pressure test date 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  Pressure Equipment Directive 2014/68/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. 1.5 Customer Service and Services Our Customer Service is at your disposal for technical advice and specialist enquiries. For contact information, see page 2. Due to their familiarity with the potential applications of the equipment, the Customer Service team is able to provide information on Techfors-S – Vessel Volume 15 L – Serial No. 129020 General Information Page 12 of 257 11 April 2019 whether the equipment can be used for a specific application or modified to handle the planned process. Experience of working with the equipment will be published semiregularly on the manufacturer’s website in the form of “application notes”. Furthermore, our colleagues are always interested in new information and experiences resulting from user’s applications for the equipment that may be valuable for the continued development of our products. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Safety and Responsibility 11 April 2019 Page 13 of 257 2 Safety and Responsibility This section describes general considerations relating to user safety that must be taken into account when working with the equipment. In the remaining sections, warning notices are used only to highlight particular hazards directly arising from the actions being described in the section in question. It is essential to read the operating manual carefully – especially this section and the warning notices in the text – and to follow the instructions therein. This section also refers to areas that are the responsibility of the provider due to certain risks arising from particular applications for which the equipment is used deliberately and with full awareness of the associated risks. 2.1 Intended Use, Incorrect Use and Misuse The in situ sterilisable pilot bioreactor Techfors-S from INFORS HT is designed especially for cultivation of microorganisms for research and development in a biotechnology labora- tory. The equipment is designed and constructed exclusively for the intended use described above. Intended use also includes following all the instructions in this operating manual, especially those relating to:  The installation site  User qualifications  Correct operation and maintenance  The use of undamaged hoses and glass vessels Any failure to observe the requirements specified in this manual shall be deemed incorrect use. Any use of the equipment outside the scope of the intended use as described above shall be deemed misuse. This also applies to applications for which the equipment is not designed, such as the use or production of explosive gases, which is not permitted because the equipment is not explosion-proof. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Safety and Responsibility Page 14 of 257 11 April 2019 For use for special applications not covered by conventional, intended use, the equipment must be modified and certified accordingly by the manufacturer. Any use of the equipment outside of a biotechnology laboratory, i.e. in any environment in which the conditions required for the safety of the users cannot be fulfilled or cannot be fulfilled to their full extent, shall also be deemed misuse. 2.2 Qualified Personnel Due to the complexity of the equipment and the potential risks arising from its operation, the equipment may only be used by qualified, specialist personnel. 2.2.1 Provider The term “provider” applies to all persons who are responsible for making the equipment and the necessary infrastructure available. These persons may also be included in the group of people known as “users”, though this is not always the case. Irrespective of whether a provider is a member of the company’s board of management or a supervisor, they bear a special level of responsibility with regard to the processes and the qualification and safety of the users. 2.2.2 User General The term “user” applies to all persons who come into contact with the equipment in any way and perform work on or with it. This primarily applies to the following activities, which can be performed by the manufacturer’s own specialists or a variety of other persons (it is not always possible to distinguish clearly between the different types of person):  Assembly, installation and commissioning  Definition and preparation of the process  Operation  Troubleshooting and remedying of faults  Maintenance and cleaning (autoclaving, if necessary)  Service work and repairs  Disassembly, disposal and recycling Techfors-S – Vessel Volume 15 L – Serial No. 129020 Safety and Responsibility 11 April 2019 Page 15 of 257 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. The qualified personnel (either internal or external) who cannot be categorised under the separate “operators” group are made up of the following groups of persons:  Electricians (electrical engineers)  Decontamination specialists  Repair specialists  Specialists in disassembly and (environmentally friendly) dis- posal  Recycling specialists 2.2.3 Operator The “operators” are a specific sub-group of users distinguished by the fact that they work with the equipment. They are the true target audience for this operating manual. Qualified technicians Only technicians who have been trained for working in a biotechnology laboratory can be considered for the role of operator. These include:  Process technicians in the fields of biotechnology and chemis- try  Biotechnologists (biotechnicians)  Chemists with a specialisation in biochemistry; chemists in the field of organic chemistry or biochemistry  Life scientists (biologists) with special education in cytology, bacteriology, molecular biology, genetics, etc.  Lab assistants (lab technicians) from various fields In order to be classed as a “sufficiently qualified technician” for the operation of the equipment, the persons in question must have received thorough training and have read and understood the operating manual. The operator must be informed in a training session provided by the provider of the tasks delegated to the operator and the potential risks of improper conduct. Tasks that go beyond the scope of operation under normal conditions may only be performed by the operator if this is specified in the manual and the provider has explicitly entrusted said tasks to the operator. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Safety and Responsibility Page 16 of 257 11 April 2019 Technicians in training Persons in this group who are undergoing training or apprenticeships are only permitted to use the equipment under supervision and in accordance with the instructions of a trained and qualified technician. 2.3 Unauthorised Persons The term “unauthorised persons” applies to all persons who can access the work area but are not qualified to use the equipment in accordance with the aforementioned requirements. Unauthorised persons are not permitted to operate the equipment or use it in any other way. 2.4 Responsibility of the Provider The equipment is used for industrial and scientific purposes. As such, the provider of the equipment is individually liable with regard to the legal requirements relating to occupational health and safety in a biotechnology laboratory. In particular:  The provider is responsible for ensuring that the work and environmental regulations applicable in a biotechnology laboratory are observed.  The provider must ensure that the equipment remains in safe and proper working condition throughout its entire term of use.  The provider must ensure that all safety equipment is fully functional and is not disabled.  The provider must ensure that the equipment is only worked on by qualified users, and that said users receive sufficient training.  The provider must ensure that the protective equipment required for working with the equipment is provided and worn.  The provider must ensure that this operating manual remains in the immediate vicinity of the equipment throughout its entire term of use. 2.5 General Hazards This section covers general hazards and residual risks that are always present when using the equipment in accordance with normal, intended use. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Safety and Responsibility 11 April 2019 Page 17 of 257 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. The following points must be observed in order to avoid the risk of fatal injury:  In case of damage to insulation, disconnect the equipment from the mains immediately and arrange for it to be repaired.  Disconnect the equipment from the mains before commencing any work on the electrical system.  Always use qualified electricians for any work on the electrical system.  Keep moisture away from live parts. It may lead to a short cir- cuit. 2.5.2 Unauthorised Spare Parts and Accessories Incorrect or imitated spare parts and accessories as well as spare parts or accessories that have not been authorised by the manufacturer represent a significant safety risk. As such, we recommend procuring all spare parts and accessories from an authorised dealer or directly from the manufacturer. For the contact details of the manufacturer’s representatives, see page 2. 2.6 Particular Hazards This section covers particular hazards and residual risks that may arise when using the equipment for special applications in accordance with normal, intended use. Since the use of the equipment for such applications is deliberate, it is the responsibility of the operators and the provider to ensure that all personnel are protected from potential damage to health. The provider is responsible for ensuring that the appropriate protective equipment for such applications is provided, and that the necessary infrastructure is in place. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Safety and Responsibility Page 18 of 257 11 April 2019 2.6.1 Hot Surfaces In operational state with active tempering, there is a danger of burns on hot surfaces. Since applications at high temperatures are intentional, it is the responsibility of the users to ensure that they have sufficient protec- tion. The motor gets hot during operation. There is a risk of burns if it is touched. 2.6.2 Dangerous Gases The use or production of dangerous gases i.e. toxic or asphyxiant gases entails a significant health risk, especially in enclosed spaces. In order to prevent high emissions of dangerous gases, the following measures must be taken:  The gas connections on the equipment must be checked before any cultivation processes using dangerous gases are initi- ated.  The gaskets on the equipment must be checked at regular intervals and replaced if necessary.  Siphon off exit gas safely. 2.6.3 Flammable or Explosive Substances The use or production of flammable or explosive substances is not covered under “intended use” of the equipment, as the equipment is not explosion-proof. If the provider intends to use the equipment for such purposes, he must check its suitability for the planned application with the responsible local authorities. 2.6.4 Corrosive or Toxic Substances The use or production of corrosive or toxic substances entails a significant health risk. As such, special measures must be taken to protect the users for such applications. Since the equipment is used deliberately for such applications, it is the responsibility of the users to ensure that they have sufficient protection. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Safety and Responsibility 11 April 2019 Page 19 of 257 2.6.5 Bioactive Substances or Pathogenic Organisms The use or production of bioactive substances, pathogenic organisms or genetically modified cultures entails a significant health risk. As such, special measures must be taken to protect the users for such applications. Since the equipment is used deliberately for such applications, it is the responsibility of the users to ensure that they have sufficient protection. 2.6.6 Overpressure or Vacuum The operating pressure range specified on the vessel identification plate must be adhered to. When carrying out manipulations on the vessel, it has to be considered that the vessel may be under pressure or vacuum. The correct inlet pressures and the correct pressure settings of the pressure reducing valves must be ensured at all times. 2.6.7 Steam The escape of steam can lead to severe scalding and burns! All steam-carrying components must be checked for signs of damage and for correct position before use. 2.7 Warning Symbols on the Equipment The following warning symbols (stickers) are attached to the equip- ment: Position  Instrumentation cabinet  Motor Techfors-S – Vessel Volume 15 L – Serial No. 129020 Safety and Responsibility Page 20 of 257 11 April 2019  Peristaltic pump housings WARNING Illegible or missing warning symbols on the equipment will lead to the user being exposed to risks that the warning symbols in question were designed to make him or her aware of. It is the provider’s responsibility to ensure that all the stickers with warning symbols on the equipment are always intact. 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. The following must be observed if this is the case:  The equipment, the component part or accessory must be entirely decontaminated before sending to the manufacturer  The provider is therefore required to completely and truthfully fill out a declaration of decontamination, and have it signed by the person responsible.  The declaration of decontamination must be affixed on the outer packaging in which the equipment is sent back.  These forms can be obtained from the licensed dealer or the manufacturer. See address on page 2. Important notice If the return shipment is not accompanied by a signed and complete declaration of decontamination or it is not affixed to the outer packaging, the shipment will be returned unopened to the sender at their expense (see also T&C). Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 21 of 257 3 Setup and Function 3.1 Equipment - Overview 1 Instrumentation cabinet 2 Central column 3 Vessel 4 Motor The equipment comprises the central column with support, the instrumentation cabinet with operating panel, the culture vessel, temperature control system, stirrer drive and the gassing and exit gas system. Measurement and control of pH, pO2 and antifoam is integrated by default, too. 2 3 1 4 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 22 of 257 11 April 2019 The central column with support is provided with four trolley wheels with brake. The brakes must be locked. A support bar is fitted to the central column, to secure the vessel during transportation. The hexagon screw (a) in the middle of the two threaded bolts which fixate the vessel flange is used as transport lock. A spacer (b) is located on the column for the lower end of the vessel. a) b) Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 23 of 257 3.2 Instrumentation Cabinet 1 Touch screen operating panel 2 Main switch 3 Peristaltic pumps 4 Holder for reagent bottles The instrumentation cabinet is attached to the right-hand side of the central column. It contains the entire measurement and control system. 3.2.1 Main Switch The main switch can be found at the top-right of the front side of the instrumentation cabinet. TO SWITCH ON Turn the main switch clockwise (quarter turn) in position I/ON and the power indication light is illuminated. The equipment is switched on and the bioreactor is in idle state (Phase stopped). TO SWITCH OFF Turn the main switch counter clockwise (quarter turn) in position 0/OFF. The equipment is disconnected from the power supply. Only the mains supply feed is still electrified. If necessary, secure the main switch against re-starting using a lock (not provided) and additionally unplug the power cable when starting maintenance work. 2 3 4 1 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 24 of 257 11 April 2019 ATTENTION Switching the equipment off at the main switch without previously stopping the bioreactor and/or shutting down the system on the operating panel may lead to damage of the operating panel! 3.2.2 Pumps The four peristaltic pumps for addition of reagents and feed solution are located on the lower part of the instrumentation cabinet. They are driven by stepper motors and direction of rotation is clockwise in automatic running mode. The pumps are labelled according their function: Acid, Base, AF (Antifoam) and Feed1. They have a cover made that can be folded up and down. The pumps can be manually operated (forward and reverse flow) using the rocker switches above each pump head, provided that the equipment is switched on.  Push and hold the rocker switch to the right: the pump drive shafts turns clockwise.  Push and hold the rocker switch to the left: the pump drive shaft turns counter clockwise. 3.3 Touch Screen Operating Panel The touch screen operating panel is attached to a bracket mounted on the top of the instrumentation cabinet. The bracket allows vertical swivelling of the panel so that it can be adjusted to the optimum viewing angle. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 25 of 257 The operating panel has a 12" colour-touch screen with protection IP66. The operator can access the desired options and functions of the bioreactor simply, quickly and intuitively by touching/pressing the various tabs, symbols and buttons on the screen. A detailed description is given under chapter “Operation Touch Screen Software”. 3.3.1 Monitor Keys Four monitor keys are situated on the upper left side of the touch screen operating panel. 1 ON/OFF key 2 DECREASE brightness key: to set the display illumination darker 3 INCREASE brightness key: to set the display illumination brighter 4 DISPLAY key: to switch the display on/off Special details about the ON/OFF key The touch screen operating panel is switched on and off at the main switch on the basic unit. Therefore separate switching on at the ON/OFF key is not necessary. The symbol on the key is illuminated when the operating panel is switched on. 2 3 4 1 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 26 of 257 11 April 2019 INFORMATION The ON/OFF key is locked when a bioreactor (fermentation) is running. A dialogue box with the appropriate message appears when pressing the ON/OFF key by mistake during fermentation. When pressing the ON/OFF key when the bioreactor (fermentation) is stopped, a dialogue box for shutting down the system appears 3.3.2 Operating Panel Connections Six connectors labelled with different symbols are situated on the rear side of the operating panel. 1 USB2.0 x 2: for backups and service pur- poses1 2 USB2.0 x 2: (Reserve) 3 Ethernet: for Ethernet cable 1) to connect with a network 4 COM2 (Reserve) 5 COM1: for iDDC bus cable 1) (display cable), connector is labelled with RS485 additionally 6 DC: for power supply cable 1) 1) Cable supplied with equipment 1 2 3 4 5 6 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 27 of 257 3.4 Vessel – Overview 1 Vessel top plate 2 Vessel flange 3 Vessel upper section 4 Accessory: e.g. sight glass 5 Vessel jacket 6 Standard INGOLD port 1 2 3 4 5 6 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 28 of 257 11 April 2019 3.4.1 Vessel Top Plate The vessel top plate has: 1 10 mm port with blind pocket for temperature sensor 2 Handle (2 pcs.) 3 Recessed hole, 6 pcs., for fixing the vessel top plate 4 19 mm port, 9 pcs., for mounting parts/accessories such as e.g.: safety valve, antifoam sensor, manometer, vessel light, inoculation needle(s), push valves, blind plugs etc. 5 Recessed hole, 4 pcs., for baffles 6 4 mm hole, 2 pcs., for negative lead of the antifoam sensor 7 Middle port for drive hub for the motor 8 Tri-Clamp connection flanges for: exit gas cooler (a) and inlet air filter (b) 3 a) b) 1 2 6 7 4 8 5 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 29 of 257 It is equipped with:  Diaphragm valve 805 for air inlet into sparger or head room gassing (see chapter “Gassing” for details)  4 hexagon capped nuts M8 with metal washers for fixing the baffles.  4 hexagon capped nuts M10 with metal washers for fixing the vessel top plate. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 30 of 257 11 April 2019  Blanking plugs with fitted O-ring (Ø = 2,5 x 15 mm) for the 19 mm ports  Drive hub with connection nozzle with silicone hose for lubrication of the mechanical seal  Drive shaft and 2 Rushton impellers (see chapter “Stirrer” for details)  Ringsparger (see chapter “Gassing” for details). Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 31 of 257  4 baffles with their holders and spacers on their lower end. One baffle is welded to the sparger for stabilisation (see chapter “Gassing” for details).  O-ring (vessel top plate seal) 3.4.2 Vessel Flange The vessel flange is extended on one side so that it can be hinged onto the support bar. A recessed hole is located in the middle of the flat extended part. It is for the screw that serves as transport lock for fixing the vessel to the central column. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 32 of 257 11 April 2019 The 2 opposed round studs are for correct orientation, alignment and location of the vessel top plate. 4 further flat studs are for tightening down the vessel top plate. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 33 of 257 3.4.3 Vessel Jacket The vessel jacket encloses the dished base of the vessel. The vessel jacket has:  Water outlet (upper connection)  Water inlet (lower connection)  A threaded nozzle for the steam sterilisable harvest/sample valve. For details see the chapter “Harvest/Sample Valve”. 3.4.4 Standard INGOLD Ports 3 standard INGOLD ports are readily accessible on the lower part of the front side of the vessel.  1 INGOLD standard port iD = 25 mm, horizontal / G1-1/4"  2 INGOLD standard ports, iD = 25 mm, angled (15°) / G1- 1/4" Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 34 of 257 11 April 2019 Each INGOLD standard port has a blanking plug with fixed O-ring (Ø = 3 x 20 mm). 3.4.5 Vessel Accessories The vessel accessories consist of:  Sparger, for details refer to chapter “Gassing”.  Sight glass: allows a visual control of the liquid level in the vessel. The middle of the sight glass corresponds to the working volume of the vessel.  Vessel identification plate: contains important technical vessel data and is welded to the vessel upper section. Its content is described in main chapter “General Information”, “Vessel Identification Plate”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 35 of 257  4 baffles with their holders and spacers on their lower end. The baffles are screwed into the corresponding threaded holes in the vessel top plate and fixed with a metal washer and hexagon capped nut (M8). 3.5 Harvest/Sample Valve (Bottom Valve) The harvest/sample valve (bottom valve) 201 is readily mounted to the threaded nozzle on the vessel’s base. This applies also to the steam trap and the clean steam line (pressure hose) with manual valve 203. Opening and closing the harvest/sample (bottom valve) To open: turn it to the right Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 36 of 257 11 April 2019 The valve is open, the thread is invisible. To close: turn it to the left The valve is closed, the thread is visible Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 37 of 257 Sterilisation The steam trap must be fitted to the closed valve for sterilisation by the means of a clamp. The flat gasket must be placed between the connection flanges before fixating the steam trap. Sterilisation of the harvest/sample valve is started in the touch screen software on the operating panel. For details refer to the main chapter “Operation Touch Screen Software”. Unscrewing the steam trap will expose the needle in order to pierce a septum e.g. in a sample bottle when sampling. For more details about sampling refer to the chapter “Sampling” of the main chapter “Fermentation/Cultivation”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 38 of 257 11 April 2019 3.6 Stirrer The stirrer shaft is rotated clockwise by a top drive. An air cooled servo motor is used. The range of rotation speed is from 10 up to 1500 min-1. The motor is plugged onto the drive hub and thereby coupled by fitting the notch to the bolt. After uncoupling the motor from the drive hub it can simply be inserted into the hole on the central pillar. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 39 of 257 Or when in use, into the support of the lifting device for the vessel top plate. The stirrer shaft is screwed to the drive shaft in the vessel top plate and sealed with a single mechanical seal. ATTENTION Manipulation on the mechanical seal may lead to its damage! The mechanical seal must always be lubricated! Two connection nozzles with a silicone hose are located on the drive hub for this. For details see main chapter “Cleaning and Maintenance”, chapter “Lubricating the Mechanical Seal”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 40 of 257 11 April 2019 Two height adjustable Rushton impellers with 6 blades with proportions 0.3 up to 0.4 x inner diameter of the vessel are fixated to the stirrer shaft by the means of grub screw Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 41 of 257 3.7 Temperature Control & Sterilisation 1 Solenoid valve 104 (Steam in temp. control system) 2 Safety valve Y70 temperature control circuit 3 Solenoid valve 106 (Condensate temp. control system) 4 Solenoid valve 102 (Tap water in) 5 Steam generator 6 Circulation pump P01 7 Heating element (steam via injector) 8 Solenoid valve 103 (Steam in vessel jacket) 9 Solenoid valve 101 (Tap water out) The temperature control circuit consists of a heating element with integrated steam injector for heating, an integrated circulation pump which provides water circulation into the vessel jacket. A CE approved safety valve protects the temperature control circuit against too great of an overpressure (>3 bar). Temperature control The cooling is produced by using tap/utility water that leads directly into the vessel jacket respectively into the temperature control cir- cuit. 2 3 4 1 5 6 7 8 9 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 42 of 257 11 April 2019 Heating is achieved by direct steam injection into the heating element. The tempering itself is automatic and controlled by solenoid valves in the temperature control circuit. Circuit: vessel jacket – heating element - circulation pump - vessel jacket. Temperature measurement Temperature in the vessel is measured using a platinum resistance sensor (Pt100) which is inserted into the blind pocket in the vessel top plate. Sterilisation vessel and filters Heating up and holding the sterilisation temperature is fully automatic by steam produced by the integrated steam generator. Steam is directed into the vessel jacket, where the medium is heated and sterilised. Steam raised by the vessel content sterilises the inlet air and exit gas filter. Sterilisation harvest/sample valve (bottom valve) The harvest/sample valve is separately steam sterilised with steam from the integrated steam generator, too. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 43 of 257 3.8 Gassing 1 Mass flow controller FIC801 2 Solenoid valves 810 (Air), 820 (O2), 830 (N2) 3 Inlet air filter F01 A steam sterilisable diaphragm filter of the type Novasip is used as inlet gas filter. A mixture of Air, O2 and N2 is used for sparger gassing. Gas flow control takes place via one electronic mass flow controller. The gas mixture takes place before entry into the culture vessel via three solenoid valves. The gas flow rate of the three gases is set in the Flow parameter. Settings for controlling the oxygen concentration in the gas are made in the Gasmix parameter of the touch screen software. For details refer to main chapter “Operation Touch Screen Software”, chapter “Parameters”. 2 3 1 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 44 of 257 11 April 2019 A ring sparger (a) is used for gas entry into the culture medium in the vessel. The sparger is welded to one of the four baffles (b) for better stability. The sparger is fitted with an O-ring and simply inserted into the corresponding port in the vessel top plate, where the manual diaphragm valve 805 is fitted, see following section. The baffle is screwed into the corresponding threaded hole in the vessel top plate and fixed with a metal washer and hexagon capped nut (M8) like the three other baffles. Manual Diaphragm Valve 805 The diaphragm valve 805 on the vessel top plate diverts the air into the sparger or the head space. The valve must be operated manu- ally.  Position Ster: during sterilisation the valve must be in position Ster. to lead air into the head room.  Position OP: once the sterilisation process is finished and the pre-set temperature of fermentation is reached the valve must be switched back to OP. a b Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 45 of 257 3.9 Exit Gas 1 Exit gas filter F03 2 Exit gas cooler 3 Solenoid valve 901 The exit gas leaves the vessel via the exit gas cooler, the exit gas filter and solenoid valve 901 into the atmosphere. Like for inlet gas, the same steam sterilisable membrane filter of the type Novasip is used as exit gas filter. A steam sterilisable membrane filter filters of the type Novasip are used here. Before the exit gas filter is the thermodynamic exit gas cooler, which reduces liquid losses due to evaporation Effectiveness of the exit gas cooler is based on the forced flow of the exit gas stream via a tube with internal baffles. These baffles greatly increase the surface area for condensation. The baffle tube is easily dismantled and cleaned. 1 2 3 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 46 of 257 11 April 2019 3.10 pH Control pH in the medium is measured by the pH sensor and controlled by addition of reagents (acid, base). Addition of acid and base takes place via the two peristaltic pumps Acid and Base. Reagent bottles filled with acid and base are connected to the pumps and to e.g. resterilisable push valves or inoculation needles which are mounted in 19 mm ports of the vessel top plate. Measuring System The measuring system for pH is equipped and configured for analogue sensors from the manufacturer METTLER TOLEDO. Sensor  Traditional pH sensor (potential measurement against refer- ence)  Type: 405-DPAS-SC-K8S/120  Manufacturer: METTLER TOLEDO Details on technical data, use and maintenance of the pH sensors can be found in the separate documentation of the sensor manufacturer. Read and follow instructions stated in there. Calibration As a general rule: Calibration of a pH sensor always must be carried out BEFORE sterilisation. This is executed in the touch screen software. For details refer to the main chapter “Operation Touch Screen Software”, “Calibrating the pH Sensor”. Mounting of the sensor and cable connection The pH sensor is mounted into a standard INGOLD port in the vessel. For details refer to the main chapter “Preparations before Fermentation/Cultivation”, chapter “Preparing the Sensors”, “Mounting the pH Sensor and Connecting the Cable”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 47 of 257 3.11 pO2 Control The oxygen saturation of the (culture) medium is measured by the pO2 sensor, and can be adjusted as follows: Increasing the pO2 The content of the oxygen dissolved in the medium (pO2) can be increased using the following methods:  Increasing the stirrer speed  Increasing the gas volume flow rate (air and/or oxygen)  Increasing the oxygen content in the Gasmix. These approaches can also be combined. pO2 reduction In anaerobic processes, the vessel can be gassed using nitrogen. This displaces the oxygen dissolved in the medium. For details about pO2 control (cascades) refer to the main chapter “Operation Touch Screen Software”. Measuring System The measuring system for pO2 is equipped and configured for analogue and polarographic sensors from the manufacturer METTLER TOLEDO Sensor  Traditional amperometric/polarographic pO2 sensor  Type: InPro 6820/25/080  Manufacturer: METTLER TOLEDO Polarographic pO2 sensors must be polarised at initial operation or after disconnection from the voltage source. Correct calibration is not possible otherwise. This means that the pO2 sensor must be polarised before calibration. Details on technical data, use and maintenance of the pO2 sensors can be found in the separate documentation of the sensor manufacturer. Read and follow instructions stated in there. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 48 of 257 11 April 2019 Measurement und calibration Generally speaking, the following applies: Unlike measurements such as pH, which are calibrated to absolute measurement values, the oxygen measurement is always calibrated to a relative reference point. For this purpose, the calibration is set to 100 % relative oxygen saturation, usually with air at max. stirring speed and maximum gas flow rate. The actual concentration of dissolved oxygen in mmol/L may therefore vary at 100 % saturation, depending on the process. INFORMATION Calibration is always carried out AFTER sterilisation. This is carried out in the touch screen software. Depending on the specifications defined by the user, the pO2 sensor will be calibrated either before the vessel is filled with medium or afterwards, in the prepared medium. For details about calibration refer to the main chapter “Operation Touch Screen Software”, “Calibrating the pO2 Sensor”. Mounting of the sensor and cable connection The pO2 sensor is mounted into a standard INGOLD port in the vessel. For details refer to the main chapter “Preparations before Fermentation/Cultivation”, chapter “Preparing the Sensors”, Mounting the pO2 Sensor and Connecting the Cable”. 3.12 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. This can be prevented by using different methods. The most common method is adding antifoam agent. The antifoam agent is kept in a reagent bottle that is connected to the antifoam sensor and the antifoam pump via a hose. The sensor also acts as a dosing needle and is prepared like an inoculation needle. When the antifoam sensor detects foam/liquid in the vessel, a signal is generated which leads to activation of the antifoam pump. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 49 of 257 Antifoam sensor 1 Clamping adaptor with hollow screw (a), threaded housing (b) and O-ring (c) 2 Septum collar 3 Dosing needle 4 Sensor/needle tip (sharp!) The antifoam sensor can be adjusted in its mounting depth after loosening the hollow screw in the clamping adaptor. It is provided with a septum collar and equipped with two protective caps (not visible) which are not sterilisable! The dosing needle is equipped with a transparent insulation. A fixation of the sensor in the clamping adaptor which is too tight may damage the insulation. Preparation, mounting and cable connection of the antifoam sensor are described in detail in main chapter “Preparations before Fermentation/Cultivation”, chapters “Preparing the Antifoam Sensor” and “Mounting the Antifoam Sensor and Connecting the Cable”. INFORMATION The antifoam sensor is not in situ sterilisable and must be separately autoclaved! 3 2 a b 1 4 c Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 50 of 257 11 April 2019 3.13 Transport Lock The transport lock locks the vessel in its position during transport. Furthermore, it also prevents the load cell from getting damaged, if present. The vessel flange is screwed onto the support bar on the central pillar by the means of a hexagon screw M12. Equipment with load cell may not be operated with a mounted transport lock. Therefore it is removed during installation. On equipment without load cell, complete removal is not necessary, the screw must only be slightly loosened. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function 11 April 2019 Page 51 of 257 3.14 Safety Devices Safety Valves All used safety valves are TÜV approved component parts and must be integrated into the maintenance plan. Otherwise the operational safety cannot be guaranteed. For details about technical data, use and maintenance of the safety valves refer to the separate documentation from the safety valve’s manufacturers. The safety valve (a) in the temperature control circuit protects the vessel jacket against incorrect over pressure. It is fitted and adjusted by the manufacturer and no attendance is required from the user in normal operation. The safety valve (b) on the vessel top plate protects the vessel (not the jacket!) against incorrect over pressure. It must always be fitted and intact when in operational mode. b a Techfors-S – Vessel Volume 15 L – Serial No. 129020 Setup and Function Page 52 of 257 11 April 2019 The valve can be opened by turning it counter clockwise. Normally it stays closed during sterilisation and fermentation/cultivation. For exceptions see the chapter “Mounting the Safety Valve to the Vessel” of the main chapter Preparations before Fermenta- tion/Cultivation”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Accessories, Consumables and Hoses 11 April 2019 Page 53 of 257 4 Accessories, Consumables and Hoses The bioreactors Techfors-S with serial number S-129020 is purchased with different standard accessories and consumables and further optional accessories. Thes standard items are listed in the tables below. Accessories / Consumables in Starter Kit Number Reagent bottle, 500 mL 4 Cable tie, 2,4 x 85 mm, black 100 Hexagon socket spanner WAF 17 for blanking plugs in 19 mm ports in vessel top plate 1 Septum (inoculation diapghragm) for 19 mm ports 25 Disposable diaphragm filter, PTFE 0.2 µm, Ø = 25 mm 4 Double hose nipple 1/8'' x 1/8'', PVDF 10 Other Accessories Number Safety valve vessel (see “Setup & Function”, “Safety Devices”) 1 Filter, diaphragm, complete PTFE, 2 µm, for inlet air/gas and exit gas 2 Manometer for pressure indication in vessel 1 Antifoam sensor 1 Connection kit with hoses and attachments, see “Connection Kit”. 1 These standard accessories and additional accessories delivered with the equipment is described in the following chapters. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Accessories, Consumables and Hoses Page 54 of 257 11 April 2019 4.1 Connection Kit The connection kit contains the following hose and hose attachment material. Hose type Ø mm Application Pump hose 5000 mm 3.2 x 6.4 Reagent bottle connection to peristaltic pumps Pressure hose, 6 m 6.0 x 11.9 Gas connection Pressure hose, 8 m 10.0 x 17.0 Steam connection Pressure hose, 4 m 12.5 x 21.0 Condensate Attachement Ø mm Application Hose clamp, 8 pieces 12.0 Hose connection inlet air/gas Hose clamp, 6 pieces 19.0 Hose connection water inlet & outlet and steam inlet Hose clamp, 1 piece 12 – 22 Hose connection condensate 4.2 Manometer (Pressure Indication Vessel) The manometer indicates the pressure inside the vessel. It is connected to one of the 19 mm ports on the top plate and has a range from 0 – 4 bar. 1 Pressure indication 2 O-ring D=2.5x15 EPDM How to mount the manometer is described in the main chapter “Preparations before Fermentation/Cultivation”, chapter “Mounting the Manometer”. 1 2 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Accessories, Consumables and Hoses 11 April 2019 Page 55 of 257 4.3 Reagent Bottle Reagent bottles are made of borosilicate glass and are supplied readily equipped with a filter and silicone hose. 1 Silicone hose: length = 0.25 m (connected to reagent bottle) 2 Filter 3 Cable tie (2 pieces) Appropriate pump hoses are included in the starter kit. For details about preparation of a reagent bottle see the main chapter “Preparations before Fermentation/Cultivation”, chapter “Preparing Inoculation Needles and Reagent Bottles”. 1 2 3 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Accessories, Consumables and Hoses Page 56 of 257 11 April 2019 4.4 Push Valve The sterilisable push valve allows the aseptic connection of a laboratory bottle filled with e.g. feed solution or corrective reagent to a bioreactor. 1 Push valve with fitted O-ring 2 Peristaltic pump 3 Reagent bottle Preparation and use of the sterilisable push valve basically implies the following three steps: 1) Separate sterilisation in the autoclave: The whole push valve (closed!) is sterilised along with the laboratory bottle, equipped with hose line and filter, in the auto- clave. 2) In situ Sterilisation with the vessel: The exposed part of the push valve (closed!) is in situ sterilised with the vessel. 3) Fermentation/cultivation with open push valve: The push valve is opened for fermentation. To CLOSE the push valve Turn the top section of the rotating piston counter clockwise. The bottom section of the rotating piston is retracted. The upper section sticks out from the valve body. 3 2 1 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Accessories, Consumables and Hoses 11 April 2019 Page 57 of 257 To OPEN the push valve Turn the top section of the rotating piston clockwise. The bottom section of the rotating piston is pushed downwards and the gap between top section of the rotating piston and the valve body decreases. 4.5 Sensor Insertion Housing The insertion housing of the type INFIT from the manufacturer METTLER TOLEDO serves as sensor adapter for the INGOLD standard ports. It also protects the sensor connector from humidity and mechanical damage. How to mount the sensor(s) is/are described in the main chapter Preparations before Fermentation/Cultivation”. For details about technical data and use refer to the separate documentation from the manufacturer METTLER TOLEDO. 4.6 Inoculation Needles Inoculation needles are used for feeding liquids into the vessel, which cannot be in situ sterilised. These liquids may be e.g. the inoculum or heat-sensitive reagents. If using an inoculation needle, then a septum (inoculation membrane) must be fitted along with a septum collar in the appropriate port in the vessel top plate. The inoculation needle is connected with the reagent bottle and autoclaved. The liquid, e.g. the inoculum, which shall be added into the vessel, is filled into the reagent bottle under sterile condition, shortly before addition. The septum is then pierced with the inoculation needle, which is screwed into the septum collar. The septum may be wetted e.g. with an alcohol solution that is lit up before the piercing. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Accessories, Consumables and Hoses Page 58 of 257 11 April 2019 Inoculation needles are purchased including septum collar. They have an oblique tip to facilitate the piercing. The hose connection and the very sharp needle tip are covered with non-autoclavable(!) protective caps. 4.6.1 Inoculation Needle, Single 1 Protective cap (2 pces) 2 Hose connection (OD = 6.0 mm) 3 Hollow screw 4 O-Ring D = 2.5 x 15 mm, EPDM 5 Needle (ID = 4.0 mm) 6 Septum collar For details about preparation of an inoculation needle along with a reagent bottle see the main chapter “Preparations before Fermentation/Cultivation”, chapter “Preparing Inoculation Needles and Reagent Bottles”. 1 3 2 4 5 6 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Accessories, Consumables and Hoses 11 April 2019 Page 59 of 257 4.6.2 Inoculation Needle, Double 1 Protective cap (4 pces) 2 Hose connection (2 pces., OD = 4.0 mm) 3 Hollow screw 4 O-Ring D =2.5x15 EPDM 5 Needle (2 pces., ID = 2.0 mm) 6 Septum collar For details about preparation of an inoculation needle along with a reagent bottle see the main chapter “Preparations before Fermentation/Cultivation”, chapter “Preparing Inoculation Needles and Reagent Bottles”. 3 1 2 4 5 6 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Transport and Storage Page 60 of 257 11 April 2019 5 Transport and Storage The following specifications are based on transport and storage of an unpacked equipment at the provider’s site. 5.1 Transport WARNING Improper transport, the use of incorrect auxiliary equipment and careless handling of the equipment may lead to injuries and severe property damage. The following points must be observed when transporting the equipment internally (relocation):  Transportation safety devices supplied with the equipment must be mounted before relocating the equipment in order to protect it against damage.  Work at least in pairs and, where applicable, use suitable auxiliary equipment when transporting the equipment.  Especially when using auxiliary tools, it is important to observe that the equipment’s centre of gravity is not in the middle. 5.2 Storage  Before each time they are put into storage, decontaminate, thoroughly clean and dry the vessel and all accessories 1).  Store the equipment and its components clean, dry and protected against dust, dirt and liquids.  Store the equipment and its components in a cool place with low air humidity but protected against frost.  Storage temperature: 5 °C – 55 °C  Relative air humidity, non-condensing: 10 % - 95 %.  Protect the equipment from aggressive media, direct sunlight and mechanical vibrations. 1) Maintain and store sensors produced by other manufacturers in accor dance with the separate documentation. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Installation and Initial Operation 11 April 2019 Page 61 of 257 6 Installation and Initial Operation Installation and initial operation may only be carried out by the manufacturer’s qualified personnel or personnel authorised by the manufacturer. First the temperature control circuit must be filled after installation of the equipment. It is not necessary to empty the circuit, unless maintenance work on the circuit has to be done or local decisions necessitate a change. I.e. the filling has usually to be done once. Following this, the motor cable is connected and a short test run for the basic functions of the bioreactor is executed then. This work is not described in this operating manual because it must not be carried out by the operator. Therefore, this chapter only contains the connection conditions to be complied with and the services to be provided on site by the provider. WARNING Installation and initial operation requires qualified and experienced personnel. Faulty installation may lead to perilous situations or severe loss of property. Installation and initial operation after an installation are to be carried out by the manufacturer’s associates only. 6.1 General Location Requirements for Installation The following requirements must be met for the installation of the equipment:  The figures and ranges specified in the chapters “Technical Data, Connection Values” and “Technical Data, Operating Conditions” must be observed.  The equipment must only be installed inside a laboratory or a laboratory-like environment.  The installation site must be level, sufficiently stable and able to bear loads.  There must not be any sources of electrical interference near the equipment. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Installation and Initial Operation Page 62 of 257 11 April 2019 6.2 Minimum Distances To operate and maintain the equipment it must be installed with a minimum spacing of 150 mm from walls, ceilings or other equip- ment. Furthermore it must be considered, that the lifting and removing procedure of the vessel top plate including its built-in components (drive hub, sparger, stirrer shaft etc.) must be possible in an easy and convenient way. Therefore the distance between equipment and ceiling should be calculated accordingly. 6.3 Power Supply The in-house electric power supply of the equipment (bioreactor and steam generator) must meet the following conditions: Bioreactor  230 V (± 5 %)  50 Hz  1 phase L1 + N (neutral) + PE (earth) Steam generator  400 V (± 5 %)  50 Hz  3 phases L1 + L2 + L3 + N (neutral) + PE (earth) Bioreactor & steam generator  Electric power supply must be constant.  The power supply of both, the bioreactor and the steam generator, must be individually made safe by the use of an FIswitch (or RCD – Residual Current Device) of the kind RCCB, Type B on the installation site. WARNING High leakage current! Earth connection essential before connecting supply! Detailed information about technical data, operation and maintenance of the steam generator can be found in the separate documentation from its manufacturer. Read the manual BEFORE initial Techfors-S – Vessel Volume 15 L – Serial No. 129020 Installation and Initial Operation 11 April 2019 Page 63 of 257 operation of the equipment and strictly follow instructions stated in there. 6.4 Cooling Water, Drain & Condensate The in-house cooling water supply of the equipment must meet the following conditions on site:  Constant cooling water supply at a pressure of 2.0 ± 0.5 bar  Water quality “very soft” or “soft” (CaCO3 concentration 0 mmol L-1 to 1.5 mmol L-1) is appropriate. ATTENTION Cooling liquid containing alcohol may damage components of the temperature control system! Always consult the manufacturer or the responsible dealer regarding allowed cooling liquids, if the use of alcoholic cooling liquid is foreseen. Drainage The in-house drainage of the water (condensate, water outlet temperature control circuit and total emptying of the steam generator) must meet the following conditions:  The drain must be heat resistant and must not have backpres- sure.  The drain must not be close-by the user. CAUTION Hot water and/or steam can drain out from the water and condensate outlet.  The contaminated condensate must be drained off safely and disposed off environmentally correct. WARNING Health risk and danger to the environment due to contaminated condensate! Techfors-S – Vessel Volume 15 L – Serial No. 129020 Installation and Initial Operation Page 64 of 257 11 April 2019 Hoses  Only use pressure-resistant and intact hoses.  Only use hoses with appropriate diameter, use adaptors as necessary  Secure hoses with appropriate clamps. 6.5 Gas Supply The in-house supply of the process gas (air = compressed air, O2 and N2) for the equipment must meet the following conditions:  Constant gas supply at a pressure of ≥ 2.5 bar  Gas is dry, clean and free of oil and dust (recommended prefilter 10 µm)  Recommended compressed-air quality as per DIN ISO 8573- 1: Class 1,2,3,4 WARNING Using contaminated/impure gases carries a risk of explosion which may cause severe injuries and loss of property! ATTENTION Use of oily or wet compressed air may lead to damages to the mass flow controller! Hoses  Only use pressure-resistant and intact hoses.  Only use hoses with appropriate diameters, use adaptors as necessary  Secure the hoses with appropriate clamps 6.6 Exit Gas On site, it must be ensured that:  the exit gas is safely led away by means of a suitable, gastight hose.  the exit gas line/hose is higher than the exit gas filter. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Installation and Initial Operation 11 April 2019 Page 65 of 257  the working environment and/or the laboratory/laboratory-like facility is equipped with a sufficient ventilation system, depending on the application. 6.7 Steam Supply Steam supply of the equipment is be provided by an integrated steam generator. Conditions water for steam generator  Constant water supply at a pressure of min. 3 bar  Water quality: CaCO3 concentration 0 mmol L-1 to max. 0.53497mmol L-1 Detailed information about connection conditions as well as technical data, operation and maintenance of the steam generator can be found in the separate documentation from its manufacturer. Read the manual before initial operation of the equipment and strictly follow instructions stated in there. Conditions steam (vessel, inlet air and exit gas filter)  Constant steam supply at a pressure of 2 ± 0.2 bar  Required quantity of steam: approx. ≈ 8 kg/h Conditions clean steam (periphery: harvest/sample valve)  Constant clean steam supply at a pressure of 2 ± 0.2 bar  Required quantity of steam: approx. ≈ 1 kg/h  Quality: Steam that is used for sterilisation of the harvest/sample valve (bottom valve) must be of clean steam quality and can pass through a 5 micron filter Hoses  Only use pressure-resistant and intact hoses.  Only use hoses with appropriate diameter, use adaptors as necessary  Secure hoses with appropriate clamps. CAUTION The escape of steam can lead to severe burns and scalding! Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 66 of 257 11 April 2019 7 Preparations before Fermentation / Cultivation The following chapters describe the preparatory work before starting the fermentation/cultivation process. This essentially com- prises:  Preparing the vessel and accessories:  Checking gaskets (O-rings) on component parts and on the vessel  Mounting the component parts ATTENTION Mounting/removing mounting parts including blanking plugs with tools may damage them and lead to inseparable screw connec- tions! – Only mount/remove mounting parts by hand. – Use the hexagon socket spanner purchased with the equipment for mounting (hand-tight) and loosening the 19 mm blanking plugs.  Filling the vessel  Preparing sensors and other accessories  In situ Sterilisation – General Information INFORMATION The actual sterilisation processes are described in the main chapter “Operation Touch Screen Software”. 7.1 Securing the Position of the Equipment Ensure that the equipment stands stable on its working site and cannot roll away before beginning any work. CAUTION Uncontrolled rolling of the equipment contains a risk of injury and loss of property. Always lock the brakes of the trolley wheels on the base frame! Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 67 of 257 7.2 Uncoupling the Motor Before you begin, ensure the bioreactor is stopped, the system is shut down and the equipment is switched off at the main switch and the motor has cooled down. CAUTION Risk of minor burns, if touching the motor during operation or cooling phase. CAUTION The motor is heavy! Work in a team of two when uncoupling and coupling the motor. To remove the motor from the drive hub, proceed as follows: Procedure 1. Lift the motor, lightly shake it to loosen it from the drive hub as necessary. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 68 of 257 11 April 2019 2. Plug the motor into: a) the hole on the central pillar. Or if present: b) into the support of the (optional) vessel top plate lifting device. 7.3 Removing the Exit Gas Cooler and the Filters The inlet air filter and the exit gas cooler with filter (as one assembly) must be removed before removing the vessel top plate. Both are fixed by the means of clamps to the connection flanges of the vessel top plate. The exit gas filter and the inlet air/exit gas lines are fixed with clamps, too. To remove the exit gas cooler with filter and the inlet air filter, proceed as follows: Procedure 1. Unscrew the wing nut of the clamp between exit gas filter and exit gas hose. 2. Open the clamp and remove it. a) b) Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 69 of 257 3. Remove the exit gas line from the exit gas filter. Ensure that the flat gasket does not get lost. 4. Open and remove the clamp between the exit gas cooler and the connection flange on the vessel top plate in the same way as described above. 5. Remove the exit gas cooler. Ensure that the flat gasket between the flanges does not get lost. 6. Turn the exit gas cooler upside down and hinge it onto the holding device of the vessel holder. 7. Open and remove the clamp between the inlet air filter and connection flange on the vessel top plate in the same way as described for the exit gas cooler. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 70 of 257 11 April 2019 8. Remove the inlet air filter. Ensure that the flat gasket does not get lost. The inlet air filter does not need a special holder due to its light weight. It can hang loose within the connecting hoses. 9. Disconnect all additional hoses and cable connections (sensors etc.) between equipment and vessel top plate. 7.4 Removing the Vessel Top Plate In order to check O-rings on certain built-in parts and on the vessel top plate itself, it is necessary to remove it. Before you begin, ensure the following:  The bioreactor is stopped, the system is shut down and the equipment is switched off at the main switch, the vessel is non-pressurised  Any cable or hose connections between vessel top plate and basic unit or instrumentation cabinet are disconnected ATTENTION Cables or hoses connecting the vessel or its built-in parts with the instrumentation cabinet or frame can break, if they are not disconnected before removing the vessel top plate. The heavy vessel top plate can either be lifted manually or with an appropriate lifting device. In both cases two people are needed. CAUTION The vessel top plate is heavy. Risk of injuries due to inappropriate handling of the vessel top plate! Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 71 of 257 INFORMATION The use of the optional lifting device of the manufacturer of the equipment is recommended due to the heavy weight of the vessel top plate. Assembly, Operation and Maintenance of the lifting device are described in detail in the separate user manual. Proceed as follows: Proceudre 1. Loosen the capped nuts (M10) of the vessel fixing using the hexagon socket spanner provided with the equipment or with a wrench AF17. 2. Remove the capped nuts and the metal washers. 3. Disconnect any cable or hose connections between vessel top plate and basic unit or instrumentation cabinet. Manual procedure (2 people) 4. Carefully lift the vessel top plate horizontally out of the vessel holding it on both handles. Ensure that the baffles, the sparger or the stirrer do not come in contact with the inside of the vessel. ATTENTION Knocking built-in components against the inside of the vessel can cause micro scratching. In these circumstances, the certified vessel finish (inside) can no longer be guaranteed. 5. Carefully place the vessel top plate on its lid-side on an appropriate surface and ensure it cannot tilt over or fall down. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 72 of 257 11 April 2019 ATTENTION The stirrer shaft is delicate! A distorted stirrer shaft leads to unbalanced mass during operation which can damage the mechanical seal and the bearings in the hub of the drive shaft. 7.5 Checking Impellers, Baffles and Top Plate Seal The impellers, baffles and the top plate seal (O-ring) should be checked for tightness and correct position before refitting the vessel top plate. Stirrer shaft & impellers Note the following: ATTENTION Manipulation on the mechanical seal may lead to its damage! Proceed as follows: Procedure 1. Ensure the impellers are adjusted to the desired height on the stirrer shaft and are tight. If necessary: loosen the grub screws (M5x6, 2 pieces per impeller) on the impellers and adjust them in their position. Tighten the grub screws. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 73 of 257 Baffles 2. Ensure that all spacers are fitted onto the baffles and fit tightly. 3. Ensure that the metal washers are fitted and the capped nuts (M8) are tightened which fixate the 4 baffles Top Plate Seal (O-ring) 4. Ensure the intact O-ring (top plate seal) is placed firmly in the groove of the vessel top plate. 7.6 Filling the Vessel & Refitting the Vessel Top Plate The easiest way to fill the vessel is when the top plate is still removed. If doing so, then all vessel mounting parts such as e.g. sensors which are mounted to lateral INGOLD ports must be prepared and mounted beforehand. The vessel can be filled either with heat resistant culture medium or with water depending on the kind of used medium and on the user’s specifications. Refer to chapter “In situ Sterilisation – General Information” for more details. Liquid could also be filled in after refitting the top plate, using one of the different ports in the top plate. When ready, the heavy vessel top plate can be refitted now. It can either manually be lifted or with an appropriate lifting device. In both cases two people are needed. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 74 of 257 11 April 2019 INFORMATION The use of the optional lifting device of the manufacturer of the equipment is recommended due to the weight of the vessel top plate. Assembly, Operation and Maintenance of the lifting device are described in detail in the separate user manual.  Respect the same safety precautions as mentioned in chapter “Removing the Vessel Top Plate”.  Use the help of a second person to lift the vessel top plate manually. Manual procedure (2 people) To refit the vessel top plate, proceed as follows: Procedure 1. Lift the vessel top plate on both handles. 2. Align the vessel top plate with the centre of the vessel and slowly lower it. Ensure that the baffles, the sparger and the stirrer do not knock against the inside of the vessel. 3. The recessed holes in the vessel top plate must be in line with the flat studs on the vessel flange. 4. Fit the vessel top plate. Two engraved marks on the vessel top plate and on the flange of the vessel designate the orientation of the vessel top plate. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 75 of 257 5. Place the metal washers onto the flat studs. 6. Fit the capped nuts (M10) onto the flat studs and tighten them cross-wise with a wrench AF17. 7.7 Checking Lubrication of the Mechanical Seal The mechanical seal must always be lubricated. This can easily be checked. The silicone hose on the flange of the drive hub must be completely filled with glycerine, top it up as necessary. For details see the main chapter “Cleaning and Maintenance”, chapter “Lubricating the Mechanical Seal”. ATTENTION A mechanical seal, which has not been adequately lubricated is destroyed when running dry! Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 76 of 257 11 April 2019 7.8 Coupling the Motor CAUTION Risk of minor burns, if touching the motor during operation or cooling phase. CAUTION The motor is heavy! Work in a team of two when uncoupling and coupling the motor. To couple, the motor, proceed as follows: Procedure 1. Lift the motor out of: a) the hole of the central pillar. OR, if applicable: b) the holder on the lifting device for the vessel top plate. 2. Plug the motor onto the drive hub of the vessel top plate. Bring the slot of the motor coupling into line with the centre pin on the drive hub. Any other position is not usable. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 77 of 257 7.9 Preparing the Sensors Initial operation, maintenance and use of the sensors are described in detail in the sensor manufacturer’s documentations. These instructions are strictly to be followed! 7.9.1 Mounting / Connecting the Temperature Sensor The temperature sensor has no connector. It must simply be inserted into the blind pocket in the vessel top plate. Ensure the temperature sensor is inserted as far as it will go (feel contact metal to metal). 7.9.2 Calibrating the pH-Sensor A reliable pH measurement always needs a 2-point calibration with a high and low reference buffer. Calibration must be performed before each fermentation/cultivation process. This is described in detail in main chapter “Operation Touch Screen Software”, chapter “Calibrating the pH Sensor – Procedure”. 7.9.3 Mounting the pH Sensor and Connecting the Cable Only mount a calibrated pH sensor into the INGOLD vessel port. For details about calibration refer to the main chapter “Operation Touch Screen Software”, chapter “Calibrating the pH Sensor – Pro- cedure”. For details about technical data, use and maintenance of the sensor see the separate documentation from the manufacturer METTLER TOLEDO. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 78 of 257 11 April 2019 Mounting To mount the pH sensor, proceed as follows: Procedure 1. Carefully insert the (calibrated!) pH sensor into the sensor insertion housing provided with the sensor as far as it will go and screw it tight. The picture on the left shows the sensor insertion housing with the inserted pH sensor. 2. Loosen and remove the blanking plug from the INGOLD port. 3. Carefully screw the sensor into the INGOLD port. Ensure it is fitted in line with the port and sits neatly. Connecting the sensor cable To connect the sensor cable, proceed as follows: Procedure 1. Loosen and remove the anti-kink cable gland from the protective sleeve. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 79 of 257 2. Slip the protective sleeve over the sensor cable. INFORMATION Pictures of the sensor cable aid general understanding and can differ from the actual equipment as supplied. 3. Attach the sensor cable to the sensor. ATTENTION The sensor cable shield can be damaged by buckling or twisting. This may lead to faulty measurements. 4. Turn the ring on the cable connector and screw tightly. 5. Pull forward the protective sleeve and screw tightly. 6. Slip the slotted cable grommet over the sensor cable (delivered with the sensor insertion housing of the type In-Fit). Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 80 of 257 11 April 2019 7. Plug the slotted cable grommet into the protective sleeve. 7.9.4 Mounting the pO2 Sensor The pO2 sensor must be mounted into one of the INGOLD standard ports before sterilisation. It is polarised and calibrated after sterilisation. For details about technical data, use and maintenance of the sensor see the separate documentation from the manufacturer METTLER TOLEDO. To mount the pO2 sensor, proceed as follows: Procedure 1. Carefully remove the green protective cap from the sensor. 2. Loosen and remove the blanking plug from the INGOLD port. 3. Carefully screw the pO2 sensor into the INGOLD port. Ensure it is fitted in line with the port and sits neatly. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 81 of 257 7.9.5 Connecting the Sensor Cable To connect the cable to the pO2 sensor, proceed as follows: Procedure 1. Align the orange mark on the connector of the sensor cable with the orange mark on the sensor. ATTENTION The sensor cable shield can be damaged by buckling or twisting. This may lead to faulty measurements. 2. Plug the cable connector onto the sensor. 3. Turn the bajonet cap of the sensor cable clockwise und push gently towards the sensor. Ensure the cable is not twisted or buckled. 7.9.6 Polarising the pO2 Sensor (METTLER) Polarographic pO2 sensors must be polarised at initial operation or after disconnection from the voltage source. Correct calibration is not possible otherwise. This means that the pO2 sensor must be polarised before calibration. For polarisation, the sensor cable must simply be connected to the pO2 sensor and the equipment must be switched on at the main switch. Duration of polarisation (= polarisation time) depends on how long the pO2 sensor has been disconnected from the voltage source (= depolarisation time) As a general rule: if depolarisation time > 30 minutes, the minimum polarisation time is 360 minutes. More details about polarisation can be found in the separate documentation form the manufacturer METTLER TOLEDO. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 82 of 257 11 April 2019 7.9.7 Calibrating the pO2 Sensor Generally it applies that calibrating the pO2 sensor should be performed after sterilisation because sterilisation can change the gradient of the pO2 sensor. A 1-point calibration to 100 % is generally sufficient for an exact measurement and should be carried out again before each fermentation/cultivation. For details about calibration refer to the main chapter “Operation Touch Screen Software”, chapter “Calibrating the pO2 Sensor – Procedure”. 7.9.8 Preparing the Antifoam Sensor The antifoam sensor must be equipped with a silicone hose and a reagent bottle and wrapped up in aluminium foil or in a sterilising sheath and autoclaved like inoculation needles. The antifoam sensor should be adjusted in its mounting depth before separate sterilisation in the autoclave. It should be mounted too low rather than too high. Pulling it up during fermentation/cultivation carries a much lower risk of contamination than pushing it down. Preparation for autoclaving To prepare the antifoam sensor before autoclaving, proceed as follows: Procedure 1. Loosen and remove the blanking plug from the 19 mm port in the vessel top plate using the hexagon socket spanner provided with the equipment. 2. Remove the protective caps from the antifoam sensor. 3. Remove the septum collar from the antifoam sensor. 4. Screw the septum collar into the port. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 83 of 257 5. Insert the antifoam sensor into the septum collar. 6. Tighten the antifoam sensor. 7. Loosen the clamping screw with the hand. ATTENTION Adjusting the mounting depth of the antifoam sensor with a tight hollow screw can damage the insulation of the sensor! 8. Adjust the antifoam sensor to the desired mounting depth. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 84 of 257 11 April 2019 Ensure that the antifoam sensor head does not touch the hollow screw, otherwise a constant signal will be generated when the sensor is connected. 9. Tighten the hollow screw with the hand. 10. Loosen and remove the antifoam sensor from the septum col- lar. 11. Loosen and remove the septum collar and put aside for later use. 12. Connect the antifoam sensor with a reagent bottle filled with antifoam agent and sterilise in the autoclave. 7.9.9 Mounting the Antifoam Sensor and Connecting the Cable Proceed for the mounting (piercing through the septum) with the antifoam sensor the same way as with an inoculation needle after separate autoclaving and after in situ vessel sterilisation. Refer to chapter “Preparing Reagent Bottles and Inoculation Needles” for details. To connect the antifoam sensor cable, plug in the banana connectors as follows: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 85 of 257 Procedure 1. Insert the red banana connector into the lateral hole of the antifoam sensor. 2. Insert the black banana connector into the corresponding hole in the vessel top plate. 7.10 Mounting the Safety Valve The safety valve, which secures the vessel against excessive overpressure, must be mounted in operating state of the equipment (sterilisation and fermentation)! WARNING Possible overshoot of permitted pressure in the vessel cannot be relieved in a safe manner. This may lead to bursting or slew round of pressurised compo- nents. To mount it to the vessel, proceed as follows: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 86 of 257 11 April 2019 Procedure 1. Insert, screw in and tighten the safety valve into the port. 2. Ensure that the safety valve is closed. Close as necessary (see the following description with figures). To CLOSE Turn the valve head clock-wise, the valve head is turned upside as far as it will go, the mark CLOSED is visible below the valve head. To OPEN Turn the valve head counter clockwise, the valve head is turned downside, the thread is visible and the mark CLOSED cannot be seen anymore. Respect the following points: Usually the safety valve stays closed. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 87 of 257 To ensure that the air in the safety valve can be completely removed and replaced by steam, it is possible to manually open the safety valve during the heating phase briefly, up to 103 °C, and then close it again. This method is not recommended because sterilisation must then be attended by the operator and manually opening/closing the hot safety valve contains a risk of injury to the user. The risk of an incomplete sterilisation due to the presence of a small amount of air is virtually impossible with the type of valve supplied. 7.11 Mounting the Manometer The manometer PI903 for pressure indication in the vessel must be screwed into a 19 mm port in the vessel top plate before sterilisa- tion. ATTENTION The steel membrane of the manometer is very delicate and can be damaged when knocking it on solid objects or when rubbed! Carefully mount the manometer by hand. Proceed as follows: Procedure 1. Ensure the manometer is equipped with an intact O-ring. 2. Loosen and remove the blanking plug from the port using the hexagon socket spanner provided. 3. Carefully insert the manometer into the port and screw it tight by hand. Ensure it is mounted in line with the port. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 88 of 257 11 April 2019 7.12 Calibrating the Pumps If need be, pumps should be calibrated before beginning a fermentation/cultivation process. This provides an accurate value for the actual pumped volume of liquid delivered. The delivery rate is indicated in mL. Without calibration, only the number of revolutions is shown. The pumps must be calibrated before autoclaving the reagent bottles and before in-situ vessel sterilisation. For details of the procedure refer to the main chapter “Operation Touch Screen Software”, chapter “Calibrating the Pumps”. 7.13 Preparing Reagent Bottles and Inoculation Needles Reagent bottles, hoses and the inoculation needles are separately autoclaved. As a rule, bottles made of borosilicate glass are used. The seed bottle for subsequent aseptic filling close to the time of inoculation and the inoculation needle for inoculation are prepared the same way and autoclaved as described in the following. Reagent bottles are supplied readily equipped with a filter for pressure equalisation and a long piece of silicone hose. ATTENTION Damaged hoses and/or clogged sterile filter may lead to undesired pressure conditions in the reagent bottles. – Ensure each reagent bottle is equipped with an open pressure equalisation line with a clean and dry filter. – Only use clean, intact hoses and ensure they are firmly at- tached. The following section describes in detail how to prepare a reagent bottle for separate autoclaving and then how to connect it to the pump and the vessel for fermentation/cultivation. 7.13.1 Preparing and Autoclaving Proceed as follows: Procedure 1. Remove the lid of the reagent bottle. 2. Connect a piece of silicone hose to one of the two pipes inside the cap of the reagent bottle. The hose should not touch the bottom of the reagent bottle to avoid that the hose can adhere to the bottom and therefore liquid cannot be delivered anymore. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 89 of 257 INFORMATION Alternatively, the hose end can be angularly cut. In this case the hose end can touch the bottom of the reagent bottle then. 3. Secure the hose with a cable tie. 4. Close the reagent bottle with the lid again. Ensure the sealing between reagent bottle neck and lid sits and seals correctly. Ensure the sealing between reagent bottle neck and lid sits and seals correctly. 5. Connect a long piece of silicone hose to the same pipe but on the outside of the reagent bottle lid. The hose must be long enough to reach from the reagent bottle to the pump without tension or kinks. 6. Connect a short piece of silicone hose to the second pipe on the outside of the reagent bottle lid. 7. Connect the filter to the short piece of silicone hose. 8. Secure the hoses with cable ties. The picture on the left shows the equipped reagent bottle. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 90 of 257 11 April 2019 9. Thoroughly rinse the hose with distilled water before autoclav- ing. 10. Clearly label the reagent bottle according to its content. 11. Depending on the application: Fill the reagent bottle with reagent up to the required amount and refit the lid. Or where required, fill the reagent bottles under sterile conditions after autoclaving. ATTENTION Usage of the highly corrosive hydrochloric acid HCl as reagent leads to damage to components made of stainless steel such as e.g. component parts or the top plate. Use only non-corrosive acids, e.g. phosphoric acid, instead. 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. 12. Use hose connectors to connect a piece of pump hose to the silicone hose and then another piece of silicone hose to the pump hose end. The entire hose line must be long enough to reach from the reagent bottle via the pumps to the inoculation needle in the vessel top plate port without tension or kinks. 13. Connect the inoculation needle to the hose end. 14. Secure the hose end with a cable tie. 15. Lightly cap the filter on the reagent bottle and the inoculation needle with a little aluminium foil or fit the needle into the sterile housing. 16. Clamp off the reagent hose line with a hose clamp. 17. Autoclave the whole assembly together for e.g. 30 to 60 minutes at 121 °C. 7.13.2 Connecting to the Vessel and the Pump After autoclaving and enough time to cool down, the hoses on the reagent bottles must be connected to the pumps. The inoculation needles and the antifoam sensor are screwed into the ports in the vessel top plate AFTER in situ sterilisation, which Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 91 of 257 were equipped with septum collars and septa BEFORE the in situ vessel sterilisation. Connecting the reagent bottle to the pump Proceed as follows: Procedure 1. Place the reagent bottle onto the tray below the instrumentation cabinet. 2. Fold up the pump cover. Observe the direction of rotation (clockwise) of the pumps: 3. Thread the pump hose from left to right into the clamps and around the pump head. 4. Press the left clamp inwards and insert the pump hose at the same time. 5. Release the clamp. The pump hose is fixated by the left clamp now. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 92 of 257 11 April 2019 6. Lay the pump hose around the pump head and lightly push it in. 7. Slowly turn the pump head clock-wise with one hand and guide the hose with the other hand at the same time. CAUTION Operating the pump electrically (only possible once the equipment is switched on) and inserting the pump hose at the same time can lead to bruising the fingers and damage to the pump hose! Always turn the pump head with the hand when inserting the pump hose. 8. Press the right clamp inwards and insert the pump hose at the same time. 9. Release the clamp. The pump hose is threaded in and fixated by both clamps now. 10. Fold down the pump cover. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 93 of 257 Mounting the inoculation needles and the antifoam sensor Proceed as follows: Procedure 1. Loosen and remove the blanking plug out of the septum collar in the port using the hexagon socket spanner. 2. Remove the protective sheath, bag or aluminium foil from the inoculation needle/antifoam sensor. INFORMATION A few drops of 70% ethanol can be placed on the septum surface as an additional protection against contamination. 3. Instantly pierce the needle/antifoam sensor through the sep- tum. 4. Screw the inoculation needle/antifoam sensor into the septum collar. Filling the reagent and pump hoses Proceed as follows: Procedure 1. On the switched on equipment, use the rocker switch above the pump to fill the hose until it is filled up to the port in the vessel top plate. If the reagent bottles are correctly connected:  Tip the rocker switch to the right side: the pump runs forwards, liquid is pumped into the vessel.  Tip the rocker switch to the left side: the pump runs backwards, liquid. is pumped back into the reagent bot- tle. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 94 of 257 11 April 2019 7.14 Preparing Push Valves A reagent bottle used along with a push valve must be separately autoclaved first. After autoclaving and cooling down, the reagent bottle is connected to the pump and the push valve is mounted in closed(!) position to the vessel to plate port. In situ sterilisation of the vessel will also sterilise the only part of the push valve to be exposed since sterilisation in the autoclave. INFORMATION The same safety precautions as stated in chapter “Preparing Reagent Bottles and Inoculation Needles” apply and must be observed! 7.14.1 Autoclaving To prepare a push valve along with a reagent bottle for the autoclave, proceed as follows: Procedure 1. Prepare the reagent bottle as described in chapter “Preparing Reagent Bottles and Inoculation Needles”. 2. Connect the push valve to the hose end of the reagent bottle. 3. Ensure the push valve is in closed position, close as neces- sary. 4. Secure the hose end with a cable tie. 5. Lightly cap the filter on the reagent bottle and the push valve with a little aluminium foil. 6. Clamp off the reagent hose line with a hose clamp. 7. Autoclave the whole assembly together for e.g. 30 to 60 minutes at 121 °C. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 95 of 257 7.14.2 In Situ Sterilisation After autoclaving and cooling down, the following steps must be carried out:  The push valve must be mounted (still closed!) into a 19 mm vessel top plate port.  The reagent bottle must be connected to the pump. This is described in detail in chapter “Preparing Reagent Bottles and Inoculation Needles. INFORMATION The same safety precautions as stated in chapter “Preparing Reagent Bottles and Inoculation Needles” apply here, too and must be observed! 7.14.3 Fermentation/Cultivation After the in situ vessel sterilisation, the push valve must be opened and the reagent and pump hose must be filled. Refer to chapter “Preparing Reagent Bottles and Inoculation Needles” for details. 7.15 Equipping the Ports with Septa (Piercing Membranes) and Collars If working with the piercing method for later inoculation, addition of corrective reagent, antifoam reagent and feed solution (substrate), then the ports in the vessel top plate must be equipped with septa and septum collars. This also applies to the port, where the antifoam sensor is to be mounted. Proceed as follows: Procedure 1. Loosen and remove the blanking plug using the hexagon socket spanner provided with the equipment. 2. Insert the septum into the port. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 96 of 257 11 April 2019 3. Screw the septum collar into the port by hand. 4. Screw the blanking plug with fixed O-ring into the septum collar by hand. Tighten it with the hexagon socket spanner hand-tight. 7.16 Closing Unused Ports All unused ports in the vessel top plate and on the vessel in general must be closed with blanking plugs, i.e. blanking covers before sterilisation. Blanking plug vessel top plate Procedure 1. Ensure the blanking plug is equipped with an intact O-ring. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 97 of 257 2. Screw the blanking plug into the port by hand and tighten it hand tight using the hexagon socket spanner provided with the equipment. Ensure the blanking plug is fitted in line with the port. Blanking plugs INGOLD standard ports Procedure 1. Insert the blanking plug equipped with an intact O-ring into the port. 2. Screw the blanking plug hand tight. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 98 of 257 11 April 2019 7.17 Mounting the Exit Gas Cooler and the Filters For mounting the inlet air filter and the exit gas cooler with its filter, proceed as follows: Procedure 1. Hold the flange of the inlet air filter onto the connection flange on the vessel top plate. Ensure that the flat gasket is placed between both flanges. 2. Place the clamp around both flanges, shut it and tighten the wing nut. 3. Remove the exit gas cooler from its holding device. 4. Hold the flange of the exit gas cooler onto the connection flange on the vessel top plate. Ensure that the flat gasket is placed between both flanges. 5. Place the clamp around both flanges, shut it and tighten the wing nut. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 99 of 257 6. Hold the flange of the exit gas hose onto the connection flange of the exit gas filter. Ensure that the flat gasket is placed between both flanges. 7. Place the clamp around both flanges, shut it and tighten the wing nut. 7.18 Checking the Inlet Air/Gas and Exit Gas Lines and Filters A blocked inlet air or exit gas line or clogged filters may result in poor venting/gassing of the culture or even completely interrupting it. This leads to overpressure in the vessel, which may be released in a non-sterile manner via leaky vessel connections and gaskets. A humid/wet exit gas filter may get colonised by microorganisms which are growing through the exit gas line. This may contaminate the culture. Therefore, check and ensure the following points:  The air supply is correctly installed and turned on. The air supply must be turned on throughout the vessel sterilisation process so that no vacuum occurs during the cooling phase in the vessel sterilisation process. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation Page 100 of 257 11 April 2019  The filters for inlet air and exit gas are clean and dry. They are fitted correctly.  Unused twist valves on both filters are closed. Close them by turning them clockwise as necessary.  Twist valves with hose connections to inlet air filter and exit gas filter are open. Open them by turning counter clockwise as necessary.  All hoses sit neatly and are secured with hose clamps.  All hoses are intact; they are neither kinked nor damaged. 7.19 In situ Sterilisation – General Information There must be enough liquid in the vessel for in situ vessel sterilisation in order to generate a sufficient amount of steam. The exact evaporation loss during vessel sterilisation cannot be determined. Some liquid evaporates via the inlet air and exit gas lines. This will partly be compensated by adding the inoculum. Furthermore, compensation is possible by adding water before the sterilisation (see sterilisation method without culture medium) or by adding separately sterilised medium. Basically, there are different possible sterilisation methods, but sterilisation is always carried out according to the application and user specifications. Two commonly used practices are briefly described below. Sterilising the vessel with culture medium  Fill the vessel with culture medium  Sterilise the vessel  Add sterile water to compensate loss of liquid due to evaporation and if applicable, any heat-labile components under sterile conditions Techfors-S – Vessel Volume 15 L – Serial No. 129020 Preparations before Fermentation / Cultivation 11 April 2019 Page 101 of 257 Sterilising the vessel without culture medium Also when using heat-labile medium components, or components that, if combined with the medium, are not sterilisable, then the vessel must not be sterilised empty. Proceed as follows:  Fill the vessel approx. half full with water, to generate sufficient steam in the vessel during sterilisation. Add nutrient salts, if needed.  Sterilise the vessel. Either drain off the residual water after sterilisation or take this into account when adding the culture medium.  Add the culture medium and inoculum under sterile conditions All heat-labile components are usually sterile filtrated and added by injection or with the inoculum afterwards. INFORMATION The programmed sterilisation processes are described in detail in the appropriate chapters of the main chapter “Operation Touch Screen Software”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Fermentation / Cultivation Page 102 of 257 11 April 2019 8 Fermentation / Cultivation This chapter describes the work necessary for the performance of and after the completion of a cultivation, before the vessel is in situ sterilised again, if applicable, followed by being cleaned and then prepared for another cultivation. WARNING The vessel may be under pressure during operation! Removing mounting parts or the vessel top plate lead to spurting out of liquids and/or rapid exhausting of gasses. This may cause severe chemical burns, burns or intoxication. Always ensure the vessel is unpressurised before manipulating on mounting parts or on the vessel top plate. CAUTION Danger of scalding and burns due to contact with hot surfaces! The vessel, the pipework and their components can get hot during fermentation/cultivation and lead to burns! 8.1 Preparing the Medium Before the first sampling, which usually takes place as a ‘zero sample’ before inoculation, and before the inoculation itself, the medium must be warmed to the desired temperature. If necessary, the pO2 concentration and the pH are set. The time required for this depends on the working volume. For details about operation, see the main chapter “Operation Touch Screen Software”. INFORMATION Depending on the specifications defined by the user, the pO2 sensor is calibrated either before the vessel is filled with medium or afterwards, in the prepared medium. Calibration is described in the main chapter “Touch Screen Operation”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Fermentation / Cultivation 11 April 2019 Page 103 of 257 8.2 Sampling Samples are taken from the vessel to gain material for offline analysis. The frequency and exact method of sampling can vary due to the different analyses carried out by the operator. When the sample valve has cooled down after sterilisation, a sample can be taken. CAUTION Risk of burns due to contact with the hot sample valve! Proceed as follows: Procedure 1. Unscrew the steam trap from the needle by turning the steam trap to the left. The needle is visible now. 2. Pierce the needle through the septum on the sample bottle. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Fermentation / Cultivation Page 104 of 257 11 April 2019 3. Open the harvest/sample valve by turning it to the right. The valve is open; the thread is not visible anymore. 4. Fill the sample bottle with the required amount of liquid. 5. Close the harvest/sample valve by turning it to the left. The sample valve is closed, the thread is visible. 6. Pull the needle out of the septum on the sample bottle. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Fermentation / Cultivation 11 April 2019 Page 105 of 257 7. Screw the steam trap on again. The sample valve should be sterilised again now to allow enough time to cool down before taking the next sample. 8.3 Inoculation Preparations before inoculation Ensure the following before inoculation:  Medium is filled in the vessel.  Heat labile supplements are separately sterilised and added.  Reagent bottles are connected to the vessel via the pumps and sufficiently filled with corrective reagent / feed solution for the duration of the cultivation.  The pumps are primed.  Correct operating temperature has been reached.  Correct stirrer speed is set.  Sensors are calibrated and polarised where appropriate, control is set correctly (maybe not active yet).  Clamps are removed. There exist different possibilities to add the inoculum. The precise method of inoculation depends on the internal regulations and the kind of used system. Two commonly known methods are described in the following.  Via inoculation needle and septum The inoculum drips into the culture. This method contains a high risk of contamination.  Via push valve The inoculum drips into the culture. This method requires a sterile hose connection. 8.3.1 Inoculation with Inoculation Needle Proceed as follows for inoculation: Procedure 1. Fill the inoculum under sterile conditions into the prepared container. 2. Unscrew the blanking plug from the septum collar. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Fermentation / Cultivation Page 106 of 257 11 April 2019 3. If wished, place a few drops of Ethanol (70 %) on the septum before piercing. If wished, briefly flame the septum collar. 4. Remove the sterile sheathing/cover from the inoculation nee- dle. 5. Depending on the application: briefly flame the inoculation needle. 6. Immediately pierce the inoculation needle through the septum. 7. Screw the inoculation needle into the septum collar. 8. Transfer the desired volume of inoculum into the vessel. 9. Clamp off the silicone hose. 8.3.2 Inoculation with Push Valve For this, the following work must be carried out first:  The (closed!) push valve with fitted closed piece of suitable1 hose is separately sterilised in the autoclave.  The empty container/bottle for the inoculum fitted with a closed piece of suitable hose1 is separately sterilised in the autoclave.  The (closed!) push valve with the closed piece of hose is mounted into the vessel top plate port and in situ sterilised with the vessel. 1) suitable for establishing a sterile hose connection Proceed as follows for inoculation: Procedure 1. Fill the inoculum under sterile conditions into the prepared container. 2. Establish a sterile hose connection with the push valve. 3. Open the push valve and transfer the desired volume of inoculum into the vessel, use the peristaltic pump as necessary. 4. Clamp off the silicone hose, weld it as necessary or close the push valve. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Fermentation / Cultivation 11 April 2019 Page 107 of 257 8.4 Harvest The culture can be harvested at the end of the fermentation/cultivation process. To prevent possible sedimentation from the culture, the stirrer can be switched on during harvest. If necessary, activate gassing for sensible cultures. However, all other parameters should be switched off, provided there are no other specifications for the user. The simplest and safest preparations for harvest are made before the fermentation/cultivation is stopped, e.g. prepare an appropriate container or connect an appropriate hose to the harvest valve. CAUTION Risk of burns due to contact with the hot harvest valve! Basically, there are two different methods:  Emptying using gravity (bioreactor stopped)  Transfer of the culture medium using pressure (bioreactor is running) 8.5 Emptying the Vessel Depending on the user specifications, the vessel can be emptied either before or after sterilisation. A previously emptied culture vessel filled only with water for sterilisation is easier to clean afterwards. For emptying the vessel, the same options as for harvesting are available. For more information, see the chapter “Harvest”. If the culture will not be used further, it must be inactivated according to the current in-house instructions (e.g. by sterilisation or by lowering the pH value), and subsequently disposed of in an environmentally sound manner according to the local regulations. 8.6 Sterilisation after Fermentation / Cultivation Depending on internal guidelines, some accessories like reagent bottles, hoses, inoculation needles etc. are again separately autoclaved followed by cleaning after completion of the fermentation/cultivation process. The vessel is in situ sterilised again, too. In particular, this is mandatory because safety related, if carrying out Techfors-S – Vessel Volume 15 L – Serial No. 129020 Fermentation / Cultivation Page 108 of 257 11 April 2019 cultivation processes with safety-critical, pathogenic or genetically modified microorganisms. Proceed as follows: Procedure 1. Use the rocker switch of each peristaltic pump for completely emptying the reagent hoses. 2. Clamp off the reagent hoses and remove them from the pumps. 3. Remove inoculation needles under sterile conditions from the vessel top plate and replace the blanking plugs before vessel in situ sterilisation. If in use: close the push valves, sterilise them with the vessel in situ followed by autoclaving them separately. INFORMATION After emptying and before autoclaving, It is recommendable to thoroughly rinse the reagent hoses with water. Depending on internal guidelines, the hoses are disposed of in an environmental acceptable manner and new hoses are used for the next fer- mentation/cultivation. 4. If applicable, dispose of residual liquid in the reagent bottles in an environmental acceptable manner. 5. Autoclave the whole assembly (reagent bottles, hoses and inoculation needles). The components will be sterile and no longer carry a risk of microbial contamination after this process is completed suc- cessfully. 6. Carry out in situ sterilisation again. INFORMATION In any case, the exact procedure must comply with the internal guidelines and may therefore differ from the procedure described here. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 109 of 257 9 Operation Touch Screen Software This chapter contains detailed descriptions of all functions of the touch screen software that are accessible to the operator. ATTENTION Changing settings in the touch screen software by unqualified personnel or personnel with insufficient training may lead to loss of property. 9.1 Screen, Menu Navigation and Control Elements Most of the figures in this manual showing the various menus, dialogue boxes and tab pages of the touchscreen software reflect the view of a user with the user authorisation level of Technician. Refer to chapter “Security – User Management”, “User Levels for further information about user levels and access authorisation. The figures are used as examples and may therefore differ from the actual equipment configuration. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 110 of 257 11 April 2019 9.1.1 Screen Areas The screen is divided into the three sections: 1 Header 2 Main are 3 Footer Header shows the name of the equipment, operating states, login-status and the time. Two opposing vertical arrows in the header signalise that an external software like e.g. eve® has access to the OPC XML DA server of the touchscreen software. They are flashing while data is trans- mitted 2 3 1 Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 111 of 257 Main area shows main menus and submenus, e.g. main menu Batch, see figure above. Inputs are made exclusively in the main area. I.e. menus and dialogue boxes are called up by pressing buttons or input fields. Footer comprises 7 tabs which provide access to the 7 main menus. The tabs are displayed with a grey background. A selected tab is shown light grey. The following main menus are available (from left to right):  Main: shows process parameters and their values, pumps and a few valves of the bioreactor.  Batch: this is where the bioreactor (fermentation/cultivation process) and all sterilisation processes are started and stopped and sensors and pumps are calibrated.  Controller: shows the parameters of the bioreactor and offers the option of changing values.  Cascade: allows to set up a serial, parallel or parallel serial (mixed) cascade control of one or several parameters.  Trends: shows trends in the parameters, time span between 15 min. and 2 days.  System: provides access to the submenus VALVES, SECURITY, SETTINGS, WIPE SCREEN and SHUTDOWN  Alarms: shows parameter alarms, user alarms and system alarms 9.1.2 Control Elements The following control elements are used in the touchscreen soft- ware: Buttons Depending on the selected main menu or submenu and access authorisations, various buttons may be visible and available. Pressing a button either opens a sub menu, or a dialogue box or a tab page. Enabled buttons are white in colour, disabled buttons are grey in colour. Example: Start/Stop of the bioreactor (fermentation/cultivation pro- cess). Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 112 of 257 11 April 2019 Buttons, which are intended as the next logical step in the procedure, are shown in orange colour. Example: Confirmation of an entry and/or starting/ending a process with OK. Dialogue boxes and tab pages A dialogue box may contain further buttons, input fields or view boxes and tabs. A dialogue box may also contain instructions, notes, warnings or general information Example: Bioreactor operation: user interaction required dialogue box with information and instruction. Example: pH properties dialogue box with tabs which lead to the different parameter options. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 113 of 257 Pressing a tab leads to access of the respective option for the selected tab page. The tab for a selected tab page is displayed with a white background. Depending on the parameter and the access authorisations there may be more or less available options for a parameter. Input fields and view boxes They are included in various menus, dialogue boxes and tab pages. They either require the inputting of a numerical or an alphanumerical value or show these values. Example: Calibrate pH sensor dialogue box with input fields for calibration points and view boxes Sensor data. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 114 of 257 11 April 2019 Numeric keypad and alphanumeric keyboard Numerical values are entered using a numeric keypad (figure on the left). Alphanumerical values are entered using an alphanumeric keyboard (figure on the right). After pressing an input field, the appropriate pad/board appears. ON / OFF switch The ON / OFF switch is used in order to switch a function on or off.  ON: the switch is in orange colour  OFF: the switch is in white colour Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 115 of 257 Example: ON/OFF switch to enable/disable Edit function in main menu Cascades Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 116 of 257 11 April 2019 9.2 Main Menus 9.2.1 Main – Overview The main menu Main charts the bioreactor and some of its valves and offers an overview of its process parameters and pumps according to its configuration. Parameters Process parameters and their current measured values are displayed as buttons. Pressing one of these buttons leads to the parameter options. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 117 of 257 Pumps All integrated peristaltic pumps of the bioreactor are displayed as buttons at the right of the screen. The following four pumps are present by default:  Acid  Base  Antifoam  Feed The Feed 2 pump is optional. The delivered volume (in mL) of a calibrated pump is continuously shown while the bioreactor is running. This numerical value is displayed on the appropriate pump button, as the example for the Feed pump shows on the left. INFORMATION For pumps which are not calibrated, the number of rotations (standard pumps), respectively its total running duration (opt. Feed 2 pump) is displayed instead. When pressing one of the pump buttons, a dialogue box appears where the number of rotations of the selected pump can be reset to zero. The pump factor calculated during pump calibration is also visible and can be changed manually here. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 118 of 257 11 April 2019 Valves  The red colour signifies a closed valve.  The green colour signifies an open valve.  The letter A signifies, that the valve is switched to automatic mode.  The letter M signifies that the valve is switched to manual mode i.e. it is “forced”. Pressing a valve button opens a dialogue box where the state of the valve can be changed by switching On, Off, or Auto for diagnosis purposes. ATTENTION All valves are set to automatic mode (Auto) ex-factory. These settings may not be changed! Exit gas cooler The Cooler button (exit gas cooler) with the valve symbol signifies the valve (118) for the water supply of the exit gas cooler. The valve automatically opens during fermentation and in the cooling phase during full sterilisation when switched to automatic mode. If this valve is manually switched off, water supply is not possible anymore, the valve remains closed! This is indicated with the word OFF in red letters instead of the valve symbol on the Cooler but- ton. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 119 of 257 9.2.2 Batch – Start Menu The bioreactor (fermentation/cultivation process) and all sterilisation processes are started and stopped in main menu Batch. If present, this applies to the sterilisation process of the optional sample valve and the resterilisable feed line, too. pH and pO2 sensors and pumps are calibrated here, too. If required, pump hoses can automatically be filled and emptied. Recipes are stored, loaded or deleted in this menu. If the optional vessel load cell is installed, the weight display is tared here. Depending on the equipment’s configuration, access rights of the operator and operating state of the bioreactor more or less functions are present and enabled. Detailed descriptions of each function and process can be found in the appropriately named chapters in this manual. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 120 of 257 11 April 2019 9.2.3 Controller – Value Display The main menu Controller shows current values, setpoint values and controller outputs for the parameters of the bioreactor. Settings for parameters can be changed here.  Parameter: lists the available parameters. Pressing the desired parameter button leads to the parameter options, see chapter “Calling up Parameter Options”.  Value: displays the actual parameter values  Units: displays the units of the parameters  Setpoint: to enter/change setpoint values of parameters INFORMATION When the bioreactor has been stopped, setpoint values in the Controller menu are overwritten with the setpoint values set in the configuration dialogue. See the chapters “Setpoint” and “Setting Setpoint Values, Switching Parameters ON / OFF” for details. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 121 of 257  Cascade: indicates, whether and how cascade control is active and which process parameters are used. Settings for a cascade are made in the main menu Cascade. A detailed description about cascade control can be found in the chapter “Cascade Control”.  Output: displays the controller output for a parameter in % when the bioreactor is running. A switched off parameter is displayed as OFF. When the bioreactor has been stopped, all its parameters are automatically switched off. Parameters can be switched on or off here whilst the bioreactor is running by pressing the controller output (button with displayed value OFF or %). This is only possible, if the automatic mode is set in the Setpoint option of the parameter concerned.  V-Bar (vertical bar): shows a graph comparing the current value, set value and alarm limits:  Grey continuous marking: set setpoint value  Yellow marking: set alarm value (lower alarm / upper alarm).  Red marking: set critical values (lower critical / upper crit- ical)  Green bar: current value is within the alarm limits  Yellow bar: current value has exceeded the upper alarm value or dropped below the lower alarm value  Red bar: current value has exceeded the upper critical value or dropped below the lower critical value  O-Bar (controller output bar): shows a graph of the current controller output (%). Parameters which are controlled on two sides (e.g. pH and temperature) are shown as a two-part bar. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 122 of 257 11 April 2019 9.2.4 Cascade The main menu Cascade provides the option of setting up a serial, parallel or mixed cascade control of a parameter. This function is mainly used for pO2 regulation. The cascade settings are made in the left-hand section of the screen and the main section presents these schematically. The individual process parameters can be added to a cascade by dragging & dropping them. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 123 of 257 9.2.5 Trends – Trend Lines The touch screen operating unit keeps the current parameter values in a buffer and continuously charts them in the main menu Trends. This data can neither be archived nor edited or exported. The main menu Trends serves to provide quick information on the progress of the cultivation only. However, the data can be archived on computer connected via network using e.g. eve®. The parameters of the bioreactor are listed on the right-hand side of the screen. The ON/OFF switch next to each parameter allows to activate/deactivate the display of its trend line in the main area of the screen. All trend lines are normalised to the value range of the respective parameter. The maximum value (= 100 % of the normalised scale) is located on the top of the diagram, the minimum value (= 0 % of the normalised scale) on the bottom. When a parameter is selected from the list, the labels on the Y axis will switch to the value range of the selected parameter. When Common is selected, the labels on the Y axis are reverted to the normalised scale. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 124 of 257 11 April 2019 The sideways spread of the diagram can be selected via the buttons below the diagram:  15 min and 30 min: 15 and 30 minutes  1 h, 6 h and 12 h: 1, 6 and 12 hour(s)  1 d and 2 d: 1 and 2 day(s) The Background button allows to change the background colour of the diagram display (white, grey and black). Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 125 of 257 9.2.6 System – System Settings The main menu System shows the following:  Serial number  Software version  IP address of the system(s)  MAC (hardware) address  Firmware version  Manufacturer’s internet address (Domain) Two buttons are situated in the upper right side of the screen:  Update: for software updates. Refer to chapter “Update – Installing Software Updates” for details about this function.  Statistics: enables viewing some statistics of the software communication with the control board, i.e. the hardware of the bioreactor. The function is only used for fault diagnosis for the technical support from the manufacturer. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 126 of 257 11 April 2019 The menu has 5 buttons which access the submenus with various functions:  VALVES: displays the status of the digital outputs.  SECURITY: for system log-in and log-off, passwords and user management  SETTINGS: for the system and basic settings of the bioreactor  WIPE SCREEN: to lock the screen for 20 seconds, e.g. for screen cleaning  SHUTDOWN: to shut down and switch off the system A detailed description of the submenus can be found in the appropriately titled chapters. 9.2.6.1 Statistics – Software Communication with Bioreactor Hardware The Statistic function (Statistics button) in main menu System enables viewing some statistics of the software communication with the control board, i.e. the hardware of the bioreactor. This function is only used for fault diagnosis for the technical support from the manufacturer. 9.2.6.2 Update – Installing a Software Update The Update function in main menu System enables to install software updates from a USB stick onto the system. INFORMATION It is recommended to create a backup copy on an USB stick of the existing system configuration using the Backup function in submenu Settings before installing the software update. ATTENTION The existing configuration is preserved in the best possible way when performing a software update. In individual cases, e.g. for customized equipment, it may be necessary to make specific adjustments after the update. Therefore, always consult the Infors service or an authorised service partner before installing an update. Proceed as follows: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 127 of 257 Procedure 1. Use the special cable provided with the equipment and connect it to the appropriate connector (see figure on the left) on the rear side of the operating panel. 2. Connect the USB stick. 3. Log on to the system on user level Administrator. 4. Call up main menu System. 5. Press Update. The Confirmation dialogue box appears with:  Information: The system will be updated.  Instruction: Press OK to confirm.  OK: to execute the update  Cancel: to cancel and close the dialogue box without changes. 6. Press OK. Progress is continuously displayed on the screen. Once the update is completed, the system is restarted with its standard settings and the message Configuration files from previous version detected! in main menu Alarms indicates that configuration data of a previous version were detected. Restoring individual settings The Restore function is used now, to restore the previous individual bioreactor setttings. Proceed as follows: Procedure 1. If applicable, connect other USB stick with the previously saved data via Backup function. 2. In main menu System, call up submenu Settings and press Restore. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 128 of 257 11 April 2019 3. Select the file “ModifiedConfiguration_number_of_previous version”. Alternatively, select desired backup data from USB stick. 4. Continue as described in chapter “Restore – Restoring Saved Data or Factory Settings”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 129 of 257 9.2.7 Alarms – Parameter Alarms, User Alarms, System Alarms The main menu Alarms lists the parameter alarms for all running bioreactors by time of occurrence. The following user alarms and system alarms are shown here, too:  Password Expiry  Difference in board configuration  System restarted after power failure.  Invalid modbus map for Parameter xy INFORMATION This alarm can only occur if modbus settings were modified. Modbus settings can only be modified on user level Service.  No water detected: no water in the temperature control system or insufficient water supply. For details refer to main chapter Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 130 of 257 11 April 2019 “Interferences”, chapter “Interferences Temperature Control System”.  No communication: no communication between controller board and operating panel. For details refer to main chapter “Interferences”.  Requested specialized configuration not installed: Error occured while restoring saved data (Restore function) or during installation of a software update (Update function). A parameter alarm is signalled by the Alarm tab flashing light red and dark alternately. The screen contains the following columns:  Description: describes the alarm  Start: shows the date and time when the alarm started.  End: shows the date and time when the parameter alarm ended.  Confirmed: indicates confirmed and not confirmed alarms. Not confirmed alarms are confirmed by pressing Confirm. Confirmed alarms are displayed with the date, time and user. 9.2.7.1 Parameter Alarms A parameter alarm occurs as soon as the current value of a parameter is outside the set alarm tolerances. A parameter alarm is triggered as soon as a value drops below the lower alarm value or exceeds the upper alarm value. The example in the figure on the left shows: pO2: lower alarm (14.3 < 15). I.e. the current value of parameter pO2 (= 14.3 %) is below the lower alarm value (= 15 %). INFORMATION The values in brackets always refer to the current value compared with the setting of the alarm value or the critical value. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 131 of 257 9.2.7.2 System-Alarm “Difference in board configuration” A backup of each control board configuration of the equipment is stored in the touch screen. If there are differences between the backup and the current configuration after a firmware update / exchange of the control board respectively the touch screen, the alarm Difference in board configuration will occur. This signifies that the configurations do not correspond with each other. To enable to select the appropriate configuration, the Synchronize differing board configuration appears and is enabled in the Controller Board Configuration section of the main menu Settings. After selection of this function (pressing the button), the menu appears with the two following options:  Use current board configuration: to replace the backup in the touch screen with the current configuration of the control board. This is appropriate after exchange of a touch screen.  Use stored board configuration: to overwrite the configuration of the control board with the configuration from the backup. This is appropriate after a firmware update, respectively replacement of a control board. The alarm disappears as soon as the function is executed. 9.2.7.3 System Alarm “No water detected in temperature control system” The filling of the temperature control system is usually done once after installation by qualified personnel from the manufacturer or licensed dealer. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 132 of 257 11 April 2019 If the water sensor does not detect liquid in the temperature control system during fermentation or full sterilisation, the system will automatically try to refill the circuit. The status message REFILLING CIRCULATION signifies this. If the filling procedure has failed, the alarm No water detected in temperature control system will appear. Temperature control will be deactivated, the circulation pump and heating are switched off. The active process continues running. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 133 of 257 9.3 Submenus 9.3.1 Valves – Digital Controller Outputs The submenu VALVES displays the digital outputs and inputs of the control board. The overview is predominantly used for fault di- agnosis. All valves and digital outputs are set to automatic mode (Auto) exfactory. These settings must not be changed! Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 134 of 257 11 April 2019 The menu display can be selected in column Category: Display (Category) All Outputs and Inputs Outputs Outputs only Inputs Inputs only The main column contains: Main column Bit / Name Channel number and designation Mode Auto Automatic switching Manual Manual switching, outputs are forced, i.e. the automatic switching is thus disabled. Set (Switching status of the digital output) OFF / ON Output is switched off / on ReadB (electronic feedback channel, which confirms the change in status. OFF / ON Readback is switched off / on If the electrical connection is faulty, it is displayed as FALSE Back: leads back to main menu System. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 135 of 257 9.3.2 Security – User Management The submenu Security is used for logging on and off the system. This is where users can also be added or deleted, passwords can be issued and access authorisations can be assigned. More or less buttons may be enabled in this menu depending on the access authorisation of the registered user:  Login/Logout: to log on to the system / to log off from the  Change own password: to change the own password  New User: to add a new user  Edit User: to edit user settings  Remove User: to delete a user.  Set Default User/Clear Default User: to define/delete automatic user login.  Advanced password settings: to define password rules for password security. The different user levels, access authorisations and functions are described in the following chapters. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 136 of 257 11 April 2019 9.3.2.1 User Levels The system has five user levels with different access authorisations. The user levels are designated and defined as Groups. Service: This user group has access to all system and bioreactor settings. This user level is accessible for the manufacturer's (INFORS HT) qualified service employees only and access for all other users is blocked. Administrators: This user group has access to basic system and bioreactor settings. New users can only be added, altered and deleted by users allocated to the user groups Service or Administrators. The default setting for the login to the system is predefined as Administrator. Technicians: This user group has limited access to system and bioreactor settings. The default setting for the login to the system is predefined as Technician. User: This user group only has restricted access to the system. The default setting for the login to the system is predefined as User. Guests: This user group has viewing authorisations only and no access authorisations. Exception: users with this user level can select trend lines in the main Trends menu to display or hide them, change the background colour for the diagram display and select the time range for the diagram display. This user level serves as protection against unauthorised access to the system or unintentional changes to settings. This user level is set automatically as long as no user is logged on to the system. This is indicated by the words Guest mode in the header on the screen. The default password setting for the user groups Administrators, Technicians and User is the same: qwertyuiop. INFORMATION Passwords should be changed and managed by the authorised person (users with user level Administrators) after commissioning for the first time. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 137 of 257 9.3.2.2 Access Authorisations of User Groups The following tables group the various functions of the touch screen software with an indication of the access authorisations of the user groups. Optional functions, are listed, too. These functions are only present with the correspondingly ordered equipment configuration. These functions are labelled with an asterisk (*) Key:  V (view) = visible, function cannot be executed Visible means that, depending on the function, the button or the menu/dialogue can only be viewed.  E (execute) = Visible and function can be accessed for execu- tion I.e. functions are executable  Empty field = cannot be viewed and function cannot be exe- cuted BIOREACTOR User Groups Guests User Technician Admin. Service Start / Stop Fermentation/Cultivation V E E E E STERILISATION User Groups Guests User Technician Admin. Service Full Sterilisation V E E E E SIP Harvest/Sample Valve V E E E E RECIPES User Groups Guests User Technician Admin. Service Load/Start V V E E E Save V V E E E Delete V V E E E PUMPS User Groups Guests User Technician Admin. Service Calibrating pump(s) V E E E E Resetting counter (Reset) V E E E E Setting pump factor manually (Pump factor) V E E E E Filling/emptying hoses automatically (Fill/Empty Pumps) V E E E E Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 138 of 257 11 April 2019 PARAMETERS User Groups Guests User Technician Admin. Service Setpoint V E E E E Setting alarm and critical values (Upper/Lower Alarm, Upper/Lower Criti- cal) V E E E E Switching parameters on and off (Output active ON/OFF) V E E E E Calibrating pH sensor (Calibrate pH) V E E E E pH sensor: changing Slope and/or Offset (Manual calibration mode) E E E E Calibrating pO2 sensor (Calibrate pO2) V E E E E Function USE AS SETPOINT in calibration menu pO2 E E E E Function USE AS SETPOINT in all calibration menus except pO2 E E Calibrate, all except pH and pO2 V E E Calibrate, manually (manual calibration mode), all except for above mentioned E E PID E E E Options E CASCADES User Groups Guests User Technician Admin. Service Setting a cascade V E E E E Setting an advanced cascade (Advanced option) E E E TREND LINES (Trends) User Groups Guests User Technician Admin. Service Change display settings E E E E E ALARMS (Alarms) User Groups Guests User Technician Admin. Service Confirming alarms V E E E E Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 139 of 257 SYSTEM User Groups Guests User Technician Admin. Service Loading software update (Update) V V V E E Viewing statistics of communication between software and bioreactor hardware (Statistics) V V E E E DIGITAL INPUTS/OUTPUTS (System / Valves) User Groups Guests User Technician Admin. Service Manually switching digital inputs and outputs V V E E E Function User Groups SECURITY user management (Security) Guests User Technician Admin. Service Logging on to the system (Login) E E E E E Logging out from the system (Logout) E E E E Changing password (Change Password) E E E E Adding a new user (New User) V V E E Deleting a user (Remove User) V V E E Changing user settings (Edit User) V V E E Setting automatic user login (Set Default User) V V E E List of all users V V SYSTEM SETTINGS (System / Set- tings) Benutzergruppen Guests User Technician Admin. Service Settings Network settings (IP Settings) V V V E E Changing date and time (Change Time) V V V E E Files Saving data (Backup) V V V E E Restoring data (Restore) V V V E E Settings in Service Menu (Service Menu) E Exporting log files (Export Logs) V E E E Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 140 of 257 11 April 2019 Controller Board Configuration Setting codes for input channels (Input channel code) V V E Assigning/changing analogue outputs (Analog Outputs Assign/Adjust) V V E Setting extended function codes for digital outputs (Extended Digital Output Function Code) V V E Synchronising different configurations of the controller board (Synchronize differing board configura- tion) E E E Modbus settings (Modbus mapping) V V E Setting function codes for digital outputs (Digital Output Function Code) V V E Settings for balances (Balance Settings) V V V E E TEMPORARY SCREEN LOCK (System / Wipe Screen) User Groups Guests User Technician Admin. Service Activating the temporary screen lock V E E E E SYSTEM SHUTDOWN User Groups Guests User Technician Admin. Service Shutting down and switching off the system V E E E E Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 141 of 257 9.3.2.3 Login – Logging on to the System To log on to the system, proceed as follows: Procedure 1. Call up the main menu System and press Security. Submenu Security appears with:  Login: Drop-down menu with users available by default with factory settings (see picture above to the left):  User  Technician  Administrator  Other: for use by INFORS HT service employees only  Default: automatic user login without entering a password if previously set using Set Default User  Password: to enter the password.  Cancel: to cancel and close the menu without changes.  Login: to log on to the system (after password entry). 2. Select the desired user, e.g. Technician. 3. Press the input field Password. The alphanumeric keyboard appears. 4. Enter the password and confirm with the OK key. The input is saved; the alphanumeric keyboard disappears. 5. Press Login. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 142 of 257 11 April 2019 The user is logged in as Technician. This is shown in the header on the screen and the different functions are listed as buttons in the Security menu. The buttons Change Password, Logout, and Close are enabled on all user levels (except for Guest). INFORMATION On user level Administrator and above, the password rules can be configured here, too. For details refer to chapter “Password Security – Defining Password Rules”. 6. Press Close. Submenu Security disappears. 9.3.2.4 Logout – Logging off the System To log off from the system, proceed as follows: Procedure 1. Call up main menu System and press Security. Submenu Security appears. 2. Press Logout. The user is logged off, menu Security appears for login. 3. Press Cancel. Menu disappears. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 143 of 257 9.3.2.5 Change Password Users of all user groups can change their own password. In order to be able to change a password, the user must be logged on to the system. Proceed as follows: Procedure 1. Call up submenu Security and press Change Password. The Change password dialogue box appears with:  Old Password: to enter the old password  New Password: to enter the new password  Confirm Password (minimum 8 characters): to confirm then new password (here: min. 8 characters). INFORMATION Depending on the password rule settings, the password must meet different conditions. Password rules are configured on user level Administrator and above. For details refer to chapter “Password Security – Defining Password Rules”.  OK: to confirm inputs, save changes. This button is not enabled until the inputs have been made.  Cancel: to cancel without saving. 2. Press the input field Old Password. The alphanumerical keyboard appears. 3. Enter the old password and confirm with the OK key. Inputs appear as dots in the input field. 4. Proceed the same way with the new password in the input fields New Password and Confirm Password. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 144 of 257 11 April 2019 All inputs are displayed as dots in the input fields. 5. Press OK. The dialogue box disappears; the new password is saved. 9.3.2.6 New User – Adding a New User To add a new user to the user list, proceed as follows: Procedure 1. Log on to the system on user level Administrator. 2. Call up the submenu Security and press New User. The New User dialogue box appears with:  Login: to enter a new user (name).  Group: to select the user group.  New Password: to enter the password.  Confirm Password (minimum 8 characters): to confirm the password with min. 8 characters. INFORMATION Depending on the password rule settings, the password must meet different conditions. Password rules are configured on user level Administrator and above. For details refer to chapter “Password Security – Defining Password Rules”.  Validity duration [days]: to enter the validity duration of the password, choose “unlimited”, 30, 100 or 365 days.  Expire: shows the selected validity duration of the pass- word.  Enable user: to activate/deactivate access authorisation of the new user. This function is switched on by default. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 145 of 257 INFORMATION The user has no access rights and no password can be defined, if this function is deactivated.  Logout if inactive: to switch on/off the automatic logout when inactive after predefined duration.  Logout after, min: to set the expiration time of the automatic logout in minutes. This input field is enabled only, when the function is switched on.  OK: to confirm entries. This button is enabled only after input  Cancel: to cancel without saving. 3. Press Login. The alphanumeric keyboard appears. 4. Enter desired new user name, e.g. TEST, and confirm with the OK key. Input appears in the Login-field 5. Select the desired user group, e.g. Technicians 6. Press Password. The alphanumeric keyboard appears. 7. Enter the desired password and confirm with the OK key. The input appears as dots in input field. 8. Repeat the same procedure with the Confirm password input field. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 146 of 257 11 April 2019 9. Select the validity duration of the password, e.g. validity. 10. Switch on access rights, if necessary. 11. If applicable, set automatic logout, if inactive: Switch the function on and enter the desired time in minutes. Once all settings are made: 12. Press OK. The dialogue box disappears, the new user is added and shown in the user selection list. 9.3.2.7 Remove User – Deleting a User To remove a user from the, proceed as follows: Procedure 1. Log on to the system on user level Administrator. 2. Call up submenu Security. 3. Select the user to be deleted in the user selection list. The fictitious user TEST is used in this example. 4. Press Remove User. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 147 of 257 The Confirmation dialogue box appears with:  Information User “TEST” will be removed from user list:  Instruction Press Yes to confirm operation.  OK: to delete the user.  Cancel: to cancel without changes. 5. Press OK. The dialogue box disappears, the user TEST is deleted from the user selection list. 9.3.2.8 Edit User – Editing User Settings Edit User is used to change the following settings for an existing user: 1) Assign new user group. See chapter “New User – Adding a New User” for details on the procedure. 2) Change password. See chapter “Change Password” for details on the procedure. 3) Automatic user log-out when screen is inactive after a predefined time in minutes has elapsed. The first user level Guests is then set automatically. To edit settings, proceed as follows: Procedure 1. Log on to the system on user level Administrator. 2. Call up submenu Security. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 148 of 257 11 April 2019 3. Select the desired user from the user selection list, the fictitious user TEST is used in this example. 4. Press Edit User. The Edit User dialogue box appears with nearly identical input fields, view boxes, ON/OFF switches and buttons as in dialogue box New User. Exceptions:  Login: this function is not enabled here.  Change …: to change the password. The two input fields New Password and Confirm Password appear after pressing this button. 5. Make required settings. 6. Press OK. Settings are adopted; the dialogue box disappears. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 149 of 257 9.3.2.9 Set / Delete Default User – Setting or Deleting an automatic User Login Set Default User is used to set an automatic user login. I.e. a user can be defined who is then automatically logged on to the system the next time it is switched on. Clear Default User is used to delete the automatic login of a user. Proceed as follows: Procedure 1. Log on to the system on user level Administrator. 2. Call up submenu Security. 3. Touch the desired user in the user selection list. The fictitious user TEST is used in this example. 4. Press Set Default User. The selected user TEST is displayed in bold letters, the Set Default User button is only visible, but not enabled anymore. Changing the automatic user login Another user can be defined here for automatic login. When selecting the desired user in the list, the Set Default User button is enabled again. The new user is adopted for automatic user login after pressing Set Default User. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 150 of 257 11 April 2019 Deleting automatic user login The automatic user login can be deleted here, too. When selecting the defined user with the automatic user login setting in the list, the Clear Default User button is visible and enabled instead of the Set Default User button. After pressing Clear Default User, the automatic user login is de- leted. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 151 of 257 9.3.2.10 Password Security – Configuring Password Rules The Advanced password settings area is visible and enabled from user level Administrator in submenu Security. Conditions for creating new user passwords can be configured here. Proceed as follows: Procedure 1. Login to the system on user level Administrator. 2. Call up main menu System and press Security. Submenu Security appears with (lower part of the menu):  Password rules: Dropdown menu with choice of four pass word rules (see figure to the top left). The password must have at least:  8 characters, containing at least 1 number, 1 capital letter and 1 lower case letter.  8 characters, containing at least 1 number, 1 capital letter and 1 lower case letter and 1 special character.  8 characters.  8 up to max. 10 characters.  Hint: shows during creation of a new password which rules must be followed.  Passwort generations: defines the number of passwords that must be newly created, before a password may be reused.  Apply: to instantly apply the rule when creating a new password. This button is enabled as soon as a rule is changed. 3. Select the desired rule to apply and the number of new passwords that must be created until reuse of an old password is allowed. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 152 of 257 11 April 2019 4. Press Apply. The rule is saved and will be shown when creating the next password. 5. Press Close. Submenu Security disappears. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 153 of 257 9.3.3 Settings – Basic Unit Settings In the submenu Settings basic settings for the equipment are made. Depending on the access authorisation, more or less buttons are visible and enabled (for details refer to the tables in chapter “Access Authorisations of User Groups”. The figure above shows the menu on user level Administrator. The menu is divided into three areas with the following functions: Settings  IP Settings: for network settings  Change Time: to set the date and time Files  Backup: to save data.  Restore: to restore and upload saved data.  Service Menu: access only for qualified Infors service or licensed dealer.  Export Logs: to export log files. Controller Board Configuration  Input Channel Code: to set codes for input channels  Analog Outputs Assign/Adjust: to assign/change analogue outputs.  Extended Digital Output Function Code: to set function codes for extended digital outputs. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 154 of 257 11 April 2019  Synchronize differing board configuration: to synchronize differing board configurations. INFORMATION This button is only visible, if the appropriate alarm (Difference in board configuration!) has been triggered and is displayed in main menu Alarms after an update of the firmware / change of a control board. For details refer to chapter “System Alarm Difference in oard configuration”.  Modbus mapping: for Modbus settings.  Digital Output Function Code: to set function codes for digital outputs.  Balance Settings: for balance settings. INFORMATION None of the functions concerning inputs and outputs, function codes and modbus mappings are described in this manual. These functions are only accessible for Infors service or Infors licensed dealers. The Back button directs back to the main menu System. 9.3.3.1 IP Settings – Network Settings IP Settings is used to establish a network connection. This can be performed either automatically or manually. INFORMATION This is only possible, if a network calble is connected. This manual does not describe how to setup a network connection. To call up the menu to make settings, proceed as follows: Procedure 1. Log on to the system on user level Administrator. 2. Call up submenu Settings. 3. Press IP-Settings. The Network Settings menu appears with: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 155 of 257  Obtain IP settings automatically: to set IP settings automatically (default setting). Condition: a DHCP1 server is available in the network.  Use the following IP settings: to use the following IP settings. Only after pressing this button, the following fields are en- abled.  IP address: shows current IP address or to enter IP address manually.  Subnet mask: displays default gateway or allows manual input.  Default gateway: shows default gateway or allows manuel input.  „….connected“: status message of the network connec- tion. INFORMATION The status message …connected indicates that correct network connection is established. If this is not the case (no signal), the message „No active LAN connection“ appears.  OK: to save inputs and close the dialogue box.  Cancel: to close the dialogue box without changes. 1 ) Dynamic Host Configuration Protocol Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 156 of 257 11 April 2019 9.3.3.2 Change Time – Changing Date and Time Change Time enables adjusting the system date and time to the local conditions. The system is set for automatic synchronisation with the time server ex-factory. I.e. the display is corresponding with the selected time zone. Alternatively, these settings can be manually adjusted. To make settings, proceed as follows: Procedure 1. Log on to the system on user level Administrator and call up the submenu Settings. 2. Press Change Time. The Change System time dialogue box appears with:  ON/OFF switch Set time and date automatically is in position ON.  Display (from left to right) for year, month, day, hours, minutes and seconds.  Dropdown menus for time zone and city: default = Europe / Zurich  Cancel: to close the dialogue box without changes.  OK: to adopt inputs and close the dialogue box. Changes with automatic adjustment Proceed as follows: Procedure 1. Select the time zone and city in the drop down menus. 2. Press OK. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 157 of 257 Settings are saved, the dialogue box disappears. Manual changes Proceed as follows: Procedure 1. Switch automatic time and date setting off. Input fields (from left to right) for year, month, day, hours, minutes and seconds appear. 2. Press the desired input field(s), enter the value(s) via appearing numeric keypad and confirm with the OK key. Inputs are adopted. 3. Press OK. Inputs are saved, the dialogue box disappears. 9.3.3.3 Backup – Saving Data The Backup function is used to save the entire settings of the touchscreen software and the control board of the bioreactor. These data can be restored using the Restore function. Note the following:  Data can be saved on the internal memory or on a USB stick.  A data backup is only executable when all running processes are stopped.. To execute a backup, proceed as follows. Only when using a USB stick, otherwise go to step 2: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 158 of 257 11 April 2019 Procedure 1. Use the special cable provided with the equipment and connect it to the appropriate connector (see figure on the left) on the rear side of the operating panel and connect the USB stick. 2. Log on to the system on user level Administrator, call up main menu System and select submenu Settings. 3. Press Backup in the Files area. The Confirmation dialogue box appears with:  Information: You will be switched to backup mode.  Instruction: Press OK to confirm  OK: to confirm switching to data backup mode.  Cancel: to close the dialogue box without changes. 4. Press OK. The menu for data backup opens with:  Select backup destination:  local: to locally save the backup.  external: to save the backup externally on a detected and connected USB stick.  Create configuration backup: to create the backup. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 159 of 257  Delete backup: to delete the backup.  Create factory settings from backup: to create factory settings from the backup.  Cancel: to leave the menu without changes.  OK: to save the backup and leave the menu. 5. Select the backup destination and press Create configuration backup to create the backup. 6. Press OK to save the backup and leave the menu. Deleting a backup Pressing Delete backup opens a dialogue box with:  Question: Are you sure to delete the selected backup?  No: to cancel and close the dialogue box without changes.  Yes: to delete the backup and close the dialogue box. If backup on USB stick: 7. Remove the USB stick and the cable. 9.3.3.4 Restore – Restoring Saved Data or Restoring Factory Settings The Restore function enables to restore data, which have previously been saved using the Backup function. Data will be uploaded to the system again. It is also possible to restore factory settings using this function. INFORMATION Factory settings usually represent the settings of the bioreactor in as-delivered condition. In case of retrofitting of the bioreactor, these settings can be updated, too. Both is exclusively carried out by Infors Service or a licensed dealer. Note the following:  Data are either restored from the internal memory or from a USB stick, see chapter “Backup – Saving Data”.  The Restore function is only executable when all running processes are stopped. To execute the Restore function, proceed as follows: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 160 of 257 11 April 2019 Only when using a USB stick, otherwise go to step 2: Procedure 1. Use the special cable provided with the equipment and connect it to the appropriate connector on the rear side of the operating panel and connect the USB stick with the saved date (Backup data). 2. Log on to the system on user level Administrator, call up main menu System and select submenu Settings. 3. Press Restore. The Confirmation dialogue box appears with:  Information: You will be switched to restore mode.  Instruction: Press OK to confirm.  OK: to confirm switching to restore mode.  Cancel: to close the dialogue box without changing the mode. 4. Press OK. The menu for data restoring appears with:  Select Configuration for restore: to select the backup data for restoring.  Select factory settings: to select factory settings.  Cancel: to leave the menu without changes.  OK: to load the selected backup and restore data. Executing the backup for data restoring Pressing Select configuration for restore changes the menu display and shows with Select backup source the choice of the possible data sources:  local: internal memory Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 161 of 257  xy (drive) / external: detected and recognised USB stick  OK: To confirm selection After selection of the data source, a dialogue box appears with:  Are you sure to restore the selected backup?  No: to cancel and close the dialogue box without changes.  Yes: to confirm restoration and start listing data for configuration comparison. After confirmation via Yes, the screen changes and lists data for configuration comparison.  ≠ signifies a difference between Backup and current configu- ration.  = No difference between Backup and current configuration.  +/- To open/close tree  Show file / Show difference: To display file / difference INFORMATION This view for showing the difference within a file is for information purposes and mainly foreseen for Infors service or licensed Infors dealers. It shows the differences between the settings of the file to restore and the currently used version in unified format (also unidiff).  Cancel: to cancel the backup process and leave the menu.  OK: to execute the backup for restoring data. 9.3.3.5 Export Logs – Exporting Log Files The Export Log functions enables to save all log files (protocol files) as well as alarms and error messages on a USB stick. Note the following:  A USB stick is needed for the export.  Export is only executable when all running processes are stopped. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 162 of 257 11 April 2019 Proceed as follows: Procedure 1. Use the special cable provided with the equipment and connect it to the appropriate connector (see figure on the left) on the rear side of the operating panel. 2. Connect the USB stick. 3. Log on to the system on user level Technician or Administra- tor. 4. In main menu System, call up submenu Settings. 5. Press Export Logs. Data export is started. Once the export is finished, the Information dialogue box appears with:  Information Log files successfully exported to: xxxxx  OK: To confirm and close the dialogue box. 6. Press OK. The dialogue box disappears. The Zip file is stored on the USB stick now. 9.3.3.6 Balance Settings This function is used to configure the connected balances. It is possible to connect up to maximum 7 balances using the equipment manufacturers switchbox. Note the following: The balance(s) must be configured as follows: Baud rate 9600, 8 bits, no parity, 2 stop bits. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 163 of 257 Proceed as follows: Procedure 1. Connect the balance(s) or switchbox 2. Log on to the system on user level Administrator. 3. In main menu System, call up submenu Settings. 4. Press Balance Settings. The menu Balance Configuration appears with:  Information The balance(s) need to be configured for 9600 baud, 8 bits, no parity, 2 stop bits.  Dropdown menu balances connected: to select number of connected balance(s)  None: no balance  Single: one balance (without Switchbox)  Infors SwitchBox  7 dropdown menus which are enabled, once one option has been selected.  Back: to save settings and return to submenu Settings. 5. Select the desired balance(s). Dropdown menu(s) for selection of the balance type(s) connected appear(s). The choice contains the following types: none (no balance), Sartorius, Mettler, Kern KB und Ohaus 6. Select the balance type(s). 7. Press Back. Settings are adopted, submenu Settings reappears. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 164 of 257 11 April 2019 9.3.4 Wipe Screen – (Temporarily) Locking the Screen The submenu Wipe Screen has one function only: It locks the screen to prevent any inputs on the screen for 20 seconds. This allows e.g. cleaning the screen for 20 seconds if required. To activate the temporary screen lock, proceed as follows: Procedure 1. In main menu System, press Wipe Screen. The screen turns white, the remaining time is displayed in seconds (Time left: xx seconds). Once the time has elapsed, the last screen reappears auto- matically. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 165 of 257 9.3.5 Shutdown – Shutting Down the System The submenu Shutdown has one function only: it shuts down and switches off the system. The system can only shut down and switch off if all bioreactors have been stopped. Proceed as follows: Procedure 1. Stop any running process by pressing Stop in the main menu Batch, if necessary. 2. Call up the main menu System. 3. Press Shutdown. The Confirmation dialogue box appears with:  Question Do you want to shutdown the system?  OK: To shut down the system  Cancel: To close the dialogue box without changes 4. Press OK. The system shuts down and switches off. INFORMATION ALWAYS shut down the system first, only then switch the equipment off at the main switch. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 166 of 257 11 April 2019 9.4 Recipes The various buttons for the Recipes function in main menu Batch can be used to load and start, save or delete what are referred to as recipes. This means all parameter settings (including cascade settings) for a fermentation process can be saved and re-used for recurring operating processes later. INFORMATION All parameter settings, cascade settings and calibration data of sensors are saved. Pump calibration data are not saved. Calibration data of sensors are not uploaded. 9.4.1 Save Recipe – Saving a Recipe Recipes can be saved when the bioreactor is running or stopped. To save a recipe, proceed as follows: Procedure 1. Log on to the system on user level Technician or above. 2. Call up main menu Batch and press Save Recipe. The Save Recipe dialogue box appears with:  Input field for the file name of the recipe being saved.  OK: to save the recipe. This button is not enabled until a file name has been en- tered.  Cancel: to cancel the process without saving. 3. Touch the input field. The alphanumeric keyboard appears. 4. Enter the desired file name and confirm with the OK key. The alphanumeric keyboard disappears Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 167 of 257 The file name is accepted, OK is enabled now. 5. Press OK. The dialogue box disappears, the recipe is saved. Recipe file name used twice If the file name for a recipe has been used twice, an Error dialogue box appears with:  The specified name for the recipe already exists  Change the name for the recipe.  OK: to close the dialogue box and enter the new name. 9.4.2 Load/Start Recipe – Loading and a Recipe All preparations for a fermentation/cultivaion process should be made before loading and starting a recipe. To load a recipe, proceed as follows: Procedure 1. Log on to the system on user level Technician or above. 2. Call up main menu Batch and press Load/Start Recipe. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 168 of 257 11 April 2019 The Load recipe dialogue box appears with:  Recipe name: lists all file names of saved recipes.  Date of change: shows date and hour of the saved rec- ipe(s).  Next: to continue. This button is not enabled until a recipe has been se- lected.  Cancel: to cancel the process without saving. 3. Select the saved recipe of bioreactor A. The selected recipe is displayed with an orange background. 4. Press Next. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 169 of 257 The Load recipe dialogue box changes views:  Parameter: lists all the available parameters.  Output Active: to switch parameters on/off.  Setpoint: lists the saved parameter setpoint values of the recipe. INFORMATION This is where setpoint values can be subsequently changed here.  OK: to start the bioreactor.  Cancel: to cancel the process without saving. 5. If applicable, change setpoints and/or switch parameters on/off. 6. Press OK. The dialogue box disappears, the bioreactor starts. 9.4.3 Delete Recipe – Deleting a Recipe Recipes can only be deleted one by one. Recipes can also be deleted during a running fermentation process. To delete a recipe, proceed as follows: Procedure 1. Log on to the system on user level Technician or above. 2. Call up main menu Batch and press Delete Recipe. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 170 of 257 11 April 2019 The Delete Recipe dialogue box appears with:  Recipe name column: file name of all saved recipes.  Changed column: date and time of the saved recipes.  OK: to delete the selected recipe. This button is not enabled until a recipe has been se- lected.  Cancel: to cancel the process without saving. 3. Select the desired recipe. The selected recipe is displayed highlighted in orange. 4. Press OK. The Confirmation dialogue box appears with:  Selected recipe will be deleted  Press OK to confirm  OK: to delete the recipe and close the dialogue box.  Cancel: to cancel the process without saving. 5. Press OK. The dialogue box disappears; the recipe is deleted. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 171 of 257 9.5 Parameters – Short Description The following sections provide a short description of the equipment’s parameters which are functional. This means, the appropriate hardware of the Techfors-S bioreactor is present and configured accordingly. 9.5.1 Temp Measures and controls the temperature in the vessel. The measured values are recorded by a platinum resistor temperature sen- sor. 9.5.2 Stirrer Measures and controls the rotation speed of the stirrer shaft. This depends on factors such as the type of vessel volume, drive system, culture viscosity and number and kind of impellers and can be found in the gassing specifications of the equipment. 9.5.3 pH Measures and controls the pH in the culture vessel in a range of pH 2 to 12. The measured values are recorded by a traditional pH sensor (potential measurement against reference) manufactured by METTLER TOLEDO The pH is controlled as standard by adding acid and base via the two digital peristaltic pumps Acid and Base. The activity of the pumps is time dependent. This means that they always operate in start/stop mode at the same speed. Control is made by a PID loop. A dead band can be used to prevent unwanted rapid dosing. Temperature compensation is a special function of the pH parameter. This function must be switched on during fermentation so that the temperature dependency of the measurement principle is cor- rected. pH of liquids is also temperature dependent which is why the pH also reacts when temperature compensation is switched on. For details about technical data, use and maintenance of the sensor see the separate documentation from the sensor manufacturer METTLER TOLEDO. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 172 of 257 11 April 2019 9.5.4 pO2 Measures the saturation of dissolved oxygen in the culture. The measuring system is configured for the use of a traditional amperometric/polarographic pO2 sensor manufactured by METTLER TOLEDO. In comparison, for example, with pH measurement which is calibrated to absolute measurements, calibrating the oxygen measurement is always performed to a relative reference point. To do this the calibration is to 100 % relative oxygen saturation, generally determined with air to a max. stirrer speed and maximum gassing rate. The actual concentration of dissolved oxygen in mmol/L can therefore differ for 100 % saturation depending on the process. The PID controller output from pO2 is generally cascaded to other parameters such as Stirrer, Flow, Feed or Gasmix. For details about technical data, use and maintenance of the sensor see the separate documentation from the manufacturer METTLER TOLEDO. 9.5.5 Antifoam Measures the fill level or the foam formation and regulates the addition of antifoam reagent via the digital peristaltic pump Antifoam in the vessel. The antifoam pump is activated as soon as the antifoam sensor comes into contact with foam / liquid. The activity of the pump is time dependent. This means that it always operates in start/stop mode at the same speed.  The Dose time must be set in seconds instead of the setpoint.  The Wait time must be set in seconds instead of setting an alarm limit. 9.5.6 Feed The addition of the feed solution (feed) is regulated via the analogue peristaltic pump Feed. Pump speed is adjustable and can be set in steps of 0.1 % within a range of 0 % to 100 %. 9.5.7 GasMix Controls the oxygen concentration in the inlet air. This is achieved by switching between air and oxygen or air and nitrogen for a 2-gas-mix system or air, oxygen and nitrogen for a 3-gas-mix sys- tem. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 173 of 257 Depending on the existing configuration this means that the relevant solenoid valve is switched on or the individual gas flow parameters are controlled. Example 2-gas-mix system with air and oxygen, supplied via a magnetic valve: Settings  cycle duration: 10 seconds (visible in the input field Eval. Time (s) in the option PID of the GasMix parameter)  setpoint in the GasMix parameter: 20 This means that:  the solenoid valve for oxygen opens for 2 seconds  the solenoid valve for process air opens for 8 seconds Setpoint 100 ≙ 10 seconds Setpoint 20 ≙ 2 seconds INFORMATION For this described configuration of the 2-gas-mix system with air + oxygen with two solenoid valves, the oxygen portion of the gas mixture cannot fall below 20.95 %. For a 3-gas-mix system the displayed value is the percentage of oxygen in the gas mixture. However, for entering the setpoint the following applies:  -100 = only nitrogen ≙ 0 % O2 content  0 = only air ≙ 21 % O2 content  100 = only oxygen ≙ 100 % O2 content Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 174 of 257 11 April 2019 If the parameter GasMix combined with the parameter GM Flow and the parameters Air Flow, O2 Flow and/or N2 Flow is installed and configured, the specified parameters are preconfigured by the equipment manufacturer in an advanced cascade for pO2 control. ATTENTION Changes to a cascade that has been preset by the equipment manufacturer may cause controller errors! 9.5.8 Flow Measures and regulates the flow of two or more process gases in the culture vessel via a single mass flow controller (thermal mass meter with integrated control valve). The measurement system is entirely electronic and the measurement is displayed according to the present configuration in L/min. If the parameter Flow is available this means that the individual process gas lines are equipped with solenoid valves, which are switched using the Gasmix parameter. Setpoint Parameter GasMix Concentrationofoxygeningasmix (displayedvalueofparameterGasMix) Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 175 of 257 9.6 Parameter Options Parameter options are setting menus for the parameters. They are shown as tab pages in the Properties dialogue box for the selected parameter. The figure above shows the example of the Temp properties dialogue box (temperature parameter). The parameters and their options (setting menus) for each individual bioreactor are called up in main menu Controller. Depending on the access authorisation and the type of the parameter, more or less options may be available. For details on user levels and their access authorisations see the chapter “Access Authorisation of User Groups”. Every properties dialogue box for each parameter has two buttons:  OK: saves inputs, closes dialogue box  Cancel: closes the dialogue box without changes Most parameters have the following options:  Setpoint: this is where setpoint values, alarm values and critical values can be set and where parameters can be switched on and off.  Calibrate: this is where the sensors’ measured values are cali- brated. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 176 of 257 11 April 2019 This option is only available for calibration of the measured values of the pH, pO2 and turbidity sensors (OPTEK system) on user levels User and Technician. All other calibration menus are only accessible on user level Administrator and above.  PID: This is where controller settings are made.  Options: This is where the basic parameter settings are made. This option is only accessible to the manufacturer’s qualified personnel. This option is not visible or enabled at any other user level. The following chapters describe the content and function of the individual tab pages, i.e. parameter menus. Each menu description is followed by either detailed setting instructions or a cross reference to the respective corresponding in these operating instruc- tions. 9.6.1 Setpoint The tab page for the Setpoint option is divided up into a three-column main area with input fields and view boxes and a Controller area. Columns  Property: designation of the input fields and view boxes  Value: values of the input fields and view boxes Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 177 of 257  Bar: graphic display of the values as in main menu Controller. For details refer to chapter “Main Menus”, “Controller – Value Display”. Input fields and view boxes  Setpoint: to enter the setpoint  Value: displays the current value  Output: Shows the controller output as a percentage.  Lower Critical: to enter the lower critical value  Lower Alarm: to enter the lower alarm value  Upper Alarm: to enter the upper alarm value  Upper Critical: to enter the upper critical value Controller  Auto: to switch on the parameter into automatic mode. In this mode, it is possible to switch the parameter on or off by touching the controller output (displayed value OFF or %) in main menu Controller during a running fermentation/cultivation.  OFF: to switch off the parameter. This mode deactivates the controller output in main menu Controller, too. The pH parameter has the additional function pH temperature comp. (pH temperature compensation). It has to be switched on during a fermentation/cultivation process so that temperature-compensated values can be generated. That means, the temperature dependency of the measuring principle will be corrected. pH of liquids is temperature-dependent, too. Therefore pH will still be responsive to temperature variation, although temperature compensation is switched on. This function also has to be switched on to calibrate the pH sensor whilst simultaneously measuring the temperature of the pH buffer solution or manually entering the temperature of the buffer solution. INFORMATION Temperature compensation is only relevant when the analogue measuring system is configured for the use of pH sensors from the manufacturer METTLER. The digital measuring system which uses pH sensors from the manufacturer HAMILTON this function is integrated into the sensor, and is therefore not used in the touch screen software. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 178 of 257 11 April 2019 9.6.1.1 Setting Setpoint Values, Switching Parameters ON / OFF Parameter setpoint values are basically set in the configuration dialogue for the bioreactor. Once the bioreactor is running, setpoint values can be changed then via main menu Controller. Parameters can be switched on or off in the configuration dialogue or via main menu Controller once the bioreactor is running, if their controller output is set to automatic mode (Auto) in the Setpoint option of the parameter. In stopped state of the bioreactor, all its parameters are automatically switched off and cannot be switched on. INFORMATION The bioreactor is always started with the settings in the configuration dialogue. Changes to these settings are saved and transferred to the next configuration dialogue. If setpoint values are changed or parameters are switched on/off whilst the bioreactor is running, these settings are only adopted for the current fermentation process. Note the following when setting setpoints: When using a lightly foaming medium, set the setpoints in parameters Stirrer (stirrer speed) and the different Flow parameter(s) as low as possible if this does not have a negative effect on the oxygen supply to the culture. If there is still heavy foaming, a chemical antifoaming agent will need to be used. In this case the Dose time and Wait time in the parameter Antifoam has to be set accordingly. Settings in the configuration dialogue To make the settings in the configuration dialogue, proceed as fol- lows: Procedure 1. Call up main menu Batch and press Start. The Configuration bioreactor operation dialogue box appears with: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 179 of 257  Parameter: lists all the available (depending on the equipment configuration) parameters  Output Active: to switch each parameter individually on or off, if the Controller in the Setpoint option of the parameter is set to automatic (Auto) mode.  Setpoint: lists all parameter setpoint values with which the bioreactor is started. Setpoint values can be changed here.  OK: to save inputs and start the bioreactor (fermenta- tion/cultivation)  Cancel: to close the dialogue box without changes 2. Press Setpoint of the desired parameter, e.g. Stirrer. The numeric keypad appears. 3. Type in the desired setpoint value, e.g. 300, and confirm with the OK key. The numeric keypad disappears; the value is accepted for the Setpoint view box/input field. 4. Repeat the same procedure for all desired parameters. 5. Switch on/off parameters as required. 6. Press OK. The dialogue box disappears, the settings are saved, and the bioreactor (fermentation/cultivation) is started. Changed settings are transferred to the next configuration dia- logue. Settings on the running bioreactor To make the settings on the running bioreactor, the following two options are available: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 180 of 257 11 April 2019 a) Directly via the Setpoint input field/view boxes and the controller output buttons in the Output column of the main menu Controller. b) In the Setpoint menu of the selected parameter in the Parameter column of the main menu Controller. INFORMATION Changed settings are adopted for the fermentation/cultivation process in progress only. Proceed as follows: Variant a) Procedure 1. Select the desired bioreactor from the selection bar, e.g. bioreactor A. 2. Call up main menu Controller. 3. Press the Setpoint input field/view box of the desired parameter, e.g. Temp. The numeric keypad appears. 4. Type in the desired setpoint value, e.g. 38, and confirm with the OK key. The numeric keypad disappears; the value is accepted for the Setpoint view box/input field. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 181 of 257 5. Press the controller output button of the parameter. The parameter is switched on, the controller output changes from OFF to the display of the corresponding numeric value in %. INFORMATION To switch the parameter i.e. controller output on or off here, is only possible, if the controller of the parameter is set to automatic (Auto) mode in its Setpoint option. See also next procedure in variant b). 6. Repeat the same procedure for all desired parameters. Variant b) Procedure 1. Call up main menu Controller.. 2. Press the desired parameter button, e.g. Temp, in the Parameter column. Temp properties menu appears with option Setpoint automatically selected. 3. Press Setpoint. The numeric keypad appears. Type in the desired setpoint value, e.g.38, and confirm with the OK key. The numeric keypad disappears; the value is accepted for the Setpoint input field. 4. Change alarm values and critical values as required. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 182 of 257 11 April 2019 For details about alarm value and critical value settings refer to chapter „Setting Alarm Values and Critical Values”. 5. Ensure the controller output is switched to automatic mode (Auto), change setting as necessary. The parameter is set switched on now. 6. Press OK. The dialogue box disappears, the settings are saved. 7. Repeat the same procedure for all desired parameters. 9.6.1.2 Setting Alarm Values and Critical Values Alarm values and critical values can be set symmetrically or asym- metrically.  Symmetrically: The difference between the setpoint value and the upper alarm value or the upper critical value = the difference between the setpoint value and the lower alarm value or the lower critical value.  Asymmetrically: The difference between the setpoint value and the upper alarm value or the upper critical value ≠ the difference between the setpoint value and the lower alarm value or the lower critical value. Upper alarm values can be set ≤ upper critical values. Lower alarm values can be set ≥ lower critical values. A parameter alarm is triggered as soon as a value drops below the lower alarm value or exceeds the upper alarm value. For details see the chapter “Alarms – Parameter Alarms, User Alarms, System Alarms”, “Parameter Alarms”. INFORMATION Alarm values and critical values have to be set by selecting the desired parameter in the main menu Controller and calling up its Setpoint option menu. The setting procedure remains the same as for setpoint values. The bioreactor can be in stopped or running state while entering these values. Invalid setpoint value or alarm limit input When an invalid setpoint, alarm or critical alarm value is entered, a corresponding Invalid input dialogue box appears after confirming the entered value via OK key. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 183 of 257 Example with input of setpoint > max. value Invalid input dialogue box appears with:  Setpoint should be <= max. value  OK: To close the dialogue box without changes and enter a new setpoint. Example with input of upper critical value > max. value Invalid input dialogue box appears with:  Your alarm limits are not ordered correctly. Upper critical should be <= max. value.  OK: To close the dialogue box without changes and enter a new value. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 184 of 257 11 April 2019 9.6.2 Calibrate The tab page for the Calibrate option contains four view boxes and a button:  Reading: shows the current measured value in digital units  Value: shows the current measured value depending on the last calibration  Slope: shows the digital value of the calculated slope of the calibration line  Offset: describes the intersection point of the calibration line with the X axis  Calibrate: to open the calibration menu The calibration menus for pH and pO2 can also be called up directly via Calibrate pH and Calibrate pO2 in the main menu Batch. These two menus are described in the following chapter in detail. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 185 of 257 9.6.3 Calibration Menus pH and pO2 Sensors The below figures show the calibration menu for METTLER pH sensors on the left-hand side and on the right, the calibration menu for METTLER pO2 sensors. Both menus look essentially the same, small differences are explained in the following menu description. The calibration menu contains the following: Header: Calibration mode  2 point: to select 2-point calibration mode. This mode is automatically selected in the calibration menu for pH sensor calibration.  1 point: to select 1-point calibration mode. If the pH sensor has to be recalibrated, this is carried out in 1point calibration mode.  Manual: to select manual calibration mode This mode is only relevant to the user for parameter pH. The menu contains: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 186 of 257 11 April 2019  Slope: to manually change the slope.  Offset: to manually correct the offset (= the same effect as recalibration in 1-point mode). For details on recalibration see the chapter “Recalibrating the pH Sensor”. INFORMATION For all other parameters the manual button is only relevant to the manufacturer’s service specialists. The necessary full menu can only be accessed at the service level. Main section: Line 1 and 3  Instruction: Please set value of the first / of the second calibration point.  Use As Setpoint (pO2): this function is only relevant and can only be used under certain circumstances at the Technician user level. For details see the chapter “Special Function USE AS SETPOINT”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 187 of 257 INFORMATION For all other parameters the USE AS SETPOINT button is only relevant to the manufacturer’s service specialists. Main section: Line 2 and 4  Instruction: Put sensor into media and confirm measure. For the pH sensor this is the buffer solution.  Sensor data: displays the measurement.  Confirm Measure: to confirm measurement. INFORMATION The input fields and buttons in line 3 and 4 are only available once the first measurement has been confirmed. Line 3 and 4 are faded out in 1-point calibration mode. Lower section  Sensor quality display bar: charts the quality of the sensor. If the green colour in the display bar is barely or not at all visi- ble:  pH sensor: replace the sensor.  pO2 sensor: replace the membrane body of the sensor in accordance with the sensor manufacturer's instructions in separate documentation. If the green colour fills less than half of the display bar:  pO2 sensor, recommended: replace the sensor in accordance with the sensor manufacturer's instructions in separate documentation.  Ref. Temp: displays the measured temperature of the reference buffer solution, or is used as an input field to manually enter the temperature of the reference buffer solution. INFORMATION This field is only relevant for the pH sensor. It is only visible and available if the temperature compensation function in the setpoint menu has been switched on. Footer  Restart calibration: to restart the calibration.  Cancel calibration: to abort calibration.  OK: to confirm calibration, close calibration menu Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 188 of 257 11 April 2019 OK and Restart are only available once at least one measurement has been confirmed. 9.6.3.1 Calibrating the pH Sensor – General Information The bioreactor system is equipped and preconfigured ex-factory with the measuring system for the use of the analogue pH sensors from the manufacturer METTLER. For all pH sensors it generally applies that a reliable pH measurement always needs a 2-point calibration with a high and low reference buffer. It also applies that the calibration should be performed again before each fermentation/cultivation process. Calibration must be performed prior to sterilisation, in other words before inserting the pH sensor in the vessel. The sequence that the two reference points are calibrated is irrelevant for the existing bioreactor measurement system. The correct temperature of the buffer solution must be measured to achieve extremely exact calibration results. This can be measured directly using the temperature sensor of the equipment during calibration. Another option is to measure the temperature externally and enter the value manually in the touch screen software. If not measured or manually input, the buffer temperature is assumed to be 20 °C. Detailed information on the calibration of the pH sensors and of the pH buffers supported by the sensors can be found in separate documentation provided by the sensor manufacturer. The following describes a 2-point calibration of a METTLER pH sensor with manual entry of buffer solution temperature. This means that the calibration takes place with temperature compensation switched on. INFORMATION This corrects the temperature dependence of the measurement principle. The pH of liquids is also temperature dependent, which is why the pH also reacts to temperature deviations when temperature compensation is switched on. 9.6.3.2 Calibrating the pH Sensor - Procedure To calibrate the pH sensor, proceed as follows: Procedure 1. Connect the sensor cable. Ensure the cable is not buckled or twisted. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 189 of 257 ATTENTION The integrity of the sensor cable can be damaged by buckling or twisting. This may lead to faulty measurements. 2. Call up main menu Controller, select parameter pH and call up Option Setpoint. 3. Switch pH temperature compensation on. 4. Press OK. The dialogue box disappears, the setting is stored. 5. Carefully remove the watering cap from the pH sensor and rinse the pH sensor with distilled water, do not rub! ATTENTION Dry wiping or rubbing a pH sensor after rinsing can cause electrostatic charge. This can greatly increase the response time and generate incorrect measurements. At most, gently dab a pH sensor after rinsing, NEVER rub or wipe it! 6. Call up main menu Batch and press Calibrate pH. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 190 of 257 11 April 2019 The calibration menu Calibrate pH sensor appears. The 2-point calibration mode is automatically selected. The Ref. Temp input field/view box is displayed. INFORMATION Without switching pH temperature compensation on before, this field is not visible. 7. Press the input field in line 1. The numeric keypad appears. 8. Type in the value of the low (or high) reference buffer confirm with the OK key. INFORMATION The order in which the reference points are calibrated is irrele- vant. The numeric keypad disappears, the value is adopted. 9. Press the Ref. Temp view box/input field. The numeric keypad appears. 10. Enter the temperature value of the buffer solution and confirm with the OK key. The numeric keypad disappears; the value is adopted. 11. Hold the pH sensor into the relevant buffer solution. The measurement (in mV) is displayed in line 2 in Sensor data. As soon as the measurement is stable: 12. Press Confirm Measure in line 2. The calibration value is accepted. The input fields and buttons in line 3 and 4 are available now. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 191 of 257 INFORMATION The signal characteristics are asymmetric. In other words, the closer the signal comes to the real value, the slower the change. The calibration is inaccurate, if the measurement is confirmed with OK before the sensor signal has completely stabilised. Wait a few minutes before confirming with OK and check the reading again, if in doubt. 13. Rinse the pH sensor with distilled water, do not rub! 14. Press the input field in line 3. The numeric keypad appears. 15. Type in the value of the high (or low) reference buffer and confirm with the OK key. INFORMATION The order in which the reference points are calibrated is irrele- vant. The numeric keypad disappears; the value is adopted. 16. Put the pH sensor into the relevant buffer solution. The measurement (in mV) is displayed in line 4 in Sensor data. As soon as the measurement is stable: 17. Press Confirm Measure in line 4. The calibration value is accepted. 18. Press OK. The dialogue box disappears, the calibration values are stored. 19. Rinse the pH sensor with distilled water, do not rub! Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 192 of 257 11 April 2019 9.6.3.3 Recalibrating a METTLER pH Sensor To compensate for a deviation (drift) in the measurement over a long-term fermentation, it is possible and sufficient to recalibrate with a 1-point calibration. This means that the pH of a sample measured using an external measurement device is accepted as the new reference value in 1-point calibration mode. The same effect is achieved by manually correcting the offset (deviation). In other words, the difference between the externally determined measurement and the displayed measurement in the culture needs to be added to or subtracted from the last calculated offset value depending on the result. The correction is made in manual calibration mode. 9.6.3.4 Polarising the pO2 Sensor (METTLER) Polarographic pO2 sensors must be polarised at initial operation or after disconnection from the voltage source. Correct calibration is not possible otherwise. This means that the pO2 sensor must be polarised before calibration. For polarisation, the sensor cable must simply be connected to the pO2 sensor and the equipment must be switched on at the main switch. Duration of polarisation (= polarisation time) depends on how long the pO2 sensor has been disconnected from the voltage source (= depolarisation time) As a general rule: if depolarisation time > 30 minutes, the minimum polarisation time is 360 minutes. More details about polarisation can be found in the separate documentation form the manufacturer METTLER TOLEDO. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 193 of 257 9.6.3.5 Calibrating the pO2 Sensor – General Information The bioreactor system is equipped and preconfigured ex-factory with the measuring system for the use of analogue, polarographic METTLER pO2 sensors. Generally it applies that: calibrating the pO2 sensor should be performed after sterilisation because sterilisation can change the gradient of the pO2 sensor. A 1-point calibration to 100 % is generally sufficient for an exact measurement and should be carried out again before each fermentation. The zero point (0 % calibration) should be checked at regular intervals of about 6 months. When performing a 2-point calibration, the 0 % calibration must always be performed before the 100 % calibration. The polarographic METTLER pO2 sensors need to be polarised before calibration. For details on this see the chapter “Polarising the pO2 Sensor”. Detailed information on the polarisation, calibration, use, maintenance and servicing can be found in the separate sensor documentation of the manufacturer. 100 % calibration conditions The 100 % calibration of the pO2 sensor is performed under the following conditions:  in the medium  at the operating temperature  at the maximum expected stirrer speed  at the maximum expected gassing rate with air or the oxygencontaining gas(es) provided for fermentation/cultivation 0 % calibration conditions The operating conditions of the 0 % calibration are the same as the 100 % calibration. One exception is the gassing. To displace oxygen out of the medium, the medium has to be gassed before and during the 0 % calibration with nitrogen instead of with air or the gas(es) used for fermentation/cultivation. 9.6.3.6 Calibrating the pO2 Sensor - Procedure The following example describes a 2-point calibration of a (polarographic analogue) METTLER pO2 sensor. This needs to take place in the correct sequence. This means that the first calibration point is 0 % (zero point), the second calibration point is 100 %. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 194 of 257 11 April 2019 Proceed as follows: Procedure 1. Connect the gassing line to nitrogen. In doing so, leave the gas supply turned off. INFORMATION Depending on the equipment configuration, there may already be a nitrogen gas line. In this case you only need to adjust the gas supply accordingly. 2. In main menu Batch, call up the bioreactor configuration menu (Start button). 3. Set the temperature setpoint and switch on the parameter. 4. Set the maximum expected stirrer speed during fermentation and switch on the parameter. 5. Start the bioreactor. 6. Slowly turn on the nitrogen supply and gas the medium. 7. Wait until the required stirrer speed and operating temperature are reached and the oxygen is displaced out of the medium. 8. Call up main menu Batch and press Calibrate pO2. The calibration menu Calibrate pO2 sensor appears. The 2-point calibration mode is automatically selected. INFORMATION The Use As Setpoint button is not relevant for this calibration. If the value in the first input field is not set to 0 (zero for zero point = 0 %): 9. Press the first input field (…first calibration point) in line 1. The numeric keypad appears. 10. Type in the value 0 confirm with the OK key. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 195 of 257 The numeric keypad disappears; the value is adopted. As soon as the measurement (line 2, Sensor data) is stable: 11. Press Confirm Measure in line 2. The value is accepted as 0 % oxygen. 12. Switch off the nitrogen gassing and switch on normal gassing. 13. Set the maximal expected gassing value during fermenta- tion/cultivation. If the value in the second input field is not set to 100 (for 100 %): 14. Press the second input field (…second calibration point) in line 3. The numeric keypad appears. 15. Type in the value 100 and confirm with the OK key. The numeric keypad disappears; the value is adopted. 16. Wait until the medium is saturated with oxygen. As soon as the measurement (line 4, Sensor data) is stable: 17. Press Confirm Measure in line 4. The value is accepted as 100 % oxygen saturation. 18. Press OK. The dialogue box disappears, the calibration is stored. Special Function USE AS SETPOINT The special function USE AS SETPOINT is for users of the Technicians user level and is only relevant in the calibration menu of the pO2 parameter when using METTLER sensors. It can only be used if a three-fold gas mixture (air/O2/N2) is installed and the parameter Gasmix is configured in a cascade for the pO2 regulation. INFORMATION For all other parameters the USE AS SETPOINT button is only relevant to the manufacturer’s service specialists. How it works In the calibration menu of the pO2 parameter (METTLER sensors): a) 0 % calibration The input 0 (%) in the input field of the first calibration point and touching the USE AS SETPOINT button causes the Gasmix parameter to switch to nitrogen for this value. b) 100 % calibration Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 196 of 257 11 April 2019 (2nd point), before entering the 100 value: The input 21 (%) in the input field of the second calibration point and touching the USE AS SETPOINT button causes the Gasmix parameter to switch to air for this value. The value can then be changed to 100 (%) in the input field and the calibration completed. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 197 of 257 9.7 PID (Control) The PID tab page is split into four horizontal areas and contains input fields for PID control settings. The following table explains the function of the individual setting values in more detail. A detailed explanation of PID control with important notes for its settings can be found in the following chapters. Note the following:  In the case of parameters which are not controlled but only measured, only the value in the Eval Time (s) input field is relevant. This value is always > 0.  If the ramp output is switched off, the value in the Ramp Size % input field is not relevant. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 198 of 257 11 April 2019 Setting value Description Prop. Term Proportional factor: The greater the discrepancy between the setpoint value and the actual value the greater the controller output. Integ. Term [1/s] The integral factor aggregates all errors over the time. If the setpoint is not achieved using the proportional factor, the integral factor adjusts the output successively until the setpoint value is achieved. An integral factor set too high will lead to oscillation of the control loop. Diff Term [s] The differential quotient calculates the change in the actual value over the time and counteracts this change to limit any overshoot. Neg. Factor The negative factor can be used to add weighting to two-sided control (+100 to -100 %) (e.g. heavy acid, light alkali). In the process 1 is the balance and 0.5 or 2 equate to the half or double the controller output accordingly. Example: Nitrogen influences the pO2 value less than oxygen, thus a negative factor of 2 can compensate for the reaction of the controller. Dead Band If a dead band is entered, no control is implemented within this value at either side of the setpoint value (symmetrically, + / -). I.e. the controller output is = 0. The dead band is used for pH control. Integ. Limit [%] The integral influence is used to ensure that the integral factor cannot increase over an indefinite period. This limits erroneous accumulation. The integral influence is set between 0 and 100 % of the controller output. Ramp output In order to perform changes slowly or step-by-step, a ramp can be introduced. This is useful above all for the stirrer speed or a mass flow valve. Ramp Size [%] The ramp size is a percentage value with which the controller output is increased step-by-step and evenly over a set time. For the set time see the value (in seconds) in the Eval Time input field. Eval Time [s] The evaluation time determines the intervals in seconds at which the PID value is recalculated. The controller speed is defined this way. A scanning time of 10 seconds is a good average value. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 199 of 257 9.7.1 Explanations of PID Control The PID function is based on a generic formula provided as exam- ple: Explanation of the formula  Error = deviation between setpoint value and actual value.  P = proportional factor, proportional response to an error, used to reach a setpoint. The bigger the value, the sharper the control.  l = integral factor, integration of the error in 1/second. A typical integral factor is < 0.05.  D = differential quotient, derivative of the error, set in seconds (mostly to 0). Be aware of the following relating to the individual factors: Proportional factor The change of the proportional factor has a considerable effect on a running process. If the proportional factor is increased excessively, this causes oscillations in the control loop around the setpoint value. Example, the pH parameter To achieve the setpoint value, a little acid, then a little base, acid again, then base etc. is added. If the proportional factor is reduced excessively, the controller hardly reacts to the deviations and never achieves the setpoint value. Integral factor The integral factor should have a low value and only be changed a little in small steps with long pauses. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 200 of 257 11 April 2019 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. The higher the entered value, the less the time (in seconds) remains for control. A higher value than 0.05 is generally of no use as it exceeds the time minimum for which the control is required. This causes fluctuations in the control circuit. Example of calculation of the integral factor The cycle duration of system oscillations is measured at 50 seconds from amplitude to amplitude. The integral factor is thus calculated as follows: 1 / (50 sec x 2) = 0.01 1/sec 1 / (50 sec x 4) = 0.005 1/sec Integral factor Seconds 0.1 10 0.05 20 0.001 100 0.005 200 Differential quotient The differential quotient is rarely required. It is set to 0 (zero) at the beginning. A high value is only necessary if major changes are made in quick succession. In all circumstances it causes the controller output to react stronger. 9.7.2 Changing the PID Controller Settings When making changes to the PID controller settings proceed as follows: Procedure 1. Make a note of the factory settings, i.e. make sure they can be restored, if necessary. 2. For readjustment of a PID controller, start with the setting for the proportional factor. Select a proportional band width as large as possible. 3. Reset the integral factor and the differential quotient to zero. 4. Increase the proportional factor until the controller causes the actual value to oscillate. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 201 of 257 5. Measure the oscillation duration. E.g. with the diagram of the parameter in the software from the equipment manufacturer. 6. Halve the proportional factor and vary the integral factor between the reciprocal value of the doubled and quadrupled oscillation duration. 9.8 Cascade Control The main menu Cascade provides the option of setting up a cascade control of a process parameter – mostly pO2. This means that the controller output parameter (=Output) of the master controller (e.g. pO2) is used as a master parameter for the slave con- troller(s). INFORMATION The master controller and slave controllers are also called master and slave. Serial cascade A deviation of the setpoint of the parameter to be controlled (master controller) influences the setpoint of the first parameter (slave controller) in the cascade. If the first parameter in the cascade reaches its maximum or minimum setpoint and the setpoint of the parameter being controlled is not yet achieved, the next parameter in the serial cascade is activated and so it continues. In the example of the left-hand figure: The parameter Stirrer, the 1st slave controller, is activated first in the cascade, to control the pO2 parameter, the master controller. The parameter AirFlow, the 2nd slave controller, is only activated when the setpoint of parameter pO2 has not been achieved by the Stirrer parameter. Parallel cascade A deviation of the setpoint of the parameter to be controlled (master controller) influences the setpoint of all parameters (slave controllers) that are in the cascade. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 202 of 257 11 April 2019 In the example of the left-hand figure: The parameters Stirrer and Air Flow, both slave controllers, are activated at the same time to control the pO2 parameter, the master controller. Parallel serial cascade A deviation of the setpoint of the parameter to be controlled (master controller) influences the setpoint of all parameters (slave controllers) that are parallel and the first element in the cascade. If the parameters that are connected in parallel reach their maximum or minimum setpoint and the setpoint of the parameter being controlled is not yet achieved, the next parameter(s) in the cascade is/are activated. In the example of the left-hand figure: The parameters Stirrer and Air Flow (master controller) are activated at the same time to control the pO2 parameter. The parameter GasMix (slave controller) is only activated when the setpoint of parameter pO2 has not been achieved by the Stirrer and AirFlow parameters. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 203 of 257 9.8.1 Setting a Cascade The different cascade settings are made in the left-hand side of the main menu Cascade. The process parameters can be merged to a cascade in the main area of the menu using drag & drop. Cascade elements (parameters) can be removed and dropped off in the recycle bin using drag&drop.  Edit: to switch on/off the edit function of the cascade. Switching off this function will also deactivate the display of the present process parameters in the main area of the menu. Once the edit function is switched on, all parameters can be merged to one or even several cascades using drag&drop. Each parameter can only be used once and in one cascade only.  Clear: to call up warning dialogue and delete cascade after confirmation.  Advanced: to switch on/off the setting mode for advanced cas- cade. INFORMATION Advanced cascades are used for customised equipment configurations. They are only set from the equipment manufacturer at the factory. Their settings and possible adjustments are equipment-specific saved at the factory. If required, they may be obtained upon request from the manufacturer. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 204 of 257 11 April 2019  Parameter name, (e.g. Stirrer): selected parameter with unit. The selected parameter visually stands out from the other parameters in the main screen area. The input fields for min./ max. and setpoint values are visible and enabled at the same time to the left-hand side.  Setp. Max. und Setp. Min.: factory settings for min. and max. setpoint values which define the adjustable value range of the selected parameter in which the cascade can change the setpoint of the cascaded parameter to control the setpoint of the master controller. These values are adjustable within this predefined value range.  Setpoint: setpoint of the parameter.  Master controller: the setpoint to be controlled.  Slave controller: the starting setpoint of the parameter from which the setpoint of the parameter of the cascade can be varied within the value range of Setp. Min. up to Setp. Max. INFORMATION In most cases, it is recommended to set the setpoint for the slave controller to the lower end of the value range (Setp. Min.)  Negative: to switch on/off the negative function of a cascade. Can be used for slave controller, if an increase of the setpoint of the slave controller leads to a decrease of the current value of the slave controller.  Output: to switch on/off the cascade and all used parameters in the cascade hereby. Each parameter in the cascade must be switched on (Output ON) for the cascade to function. The parameters can also be switched on and off in the Controller main menu. If a parameter is switched off (Output OFF), all of the following parameters are uncoupled from the cascade. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 205 of 257 Cascade progress display A cascade and its progress can be seen in the Controller main menu. In addition to arrows showing the direction of the cascade control, the setpoint and the control output of the cascade that is added to or subtracted from the setpoint is displayed in the Cascade column. These values are given in the relevant parameter unit. The colour of the added/subtracted setpoint in the Controller menu and the name of the parameter in the Cascade menu indicates the progress of the cascade and the remaining scope of the cascade within the value range of a slave controller to control the master controller according to the following scheme: Colour Utilisation of value range Grey Inactive Green 0 – 90 % Yellow 90 – 99 % Red 100 % Blue 0 % Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 206 of 257 11 April 2019 Example of calculation Stirrer, e.g. for slave controller from setpoint to max. setpoint.  Setpoint: 500  Setpoint max. 1200  Value range: 1200 – 500 = 700 700 = 100 % / 630 = 90 % 500 + 630 = 1130 = setpoint, from which 90 % of the value range are reached. This means for the display according the colour scheme men- tioned:  Green: up to 1130  Yellow: up to 1193  Red: at 1200 9.8.2 Deleting a Cascade To delete all settings of a cascade (does not apply to advanced cascade), proceed as follows: Procedure 1. In main menu Cascade, press Clear. A warning dialogue box appears with:  Warning / Instruction: All information NOT created manually with Advanced Cascades will be lost. Press OK to confirm:  OK: to confirm delete of the cascade and close the dialogue box.  Cancel: to close dialogue box without changes. 2. Press OK. Cascade is deleted. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 207 of 257 9.8.3 Negative Function of a Cascade The Negative function causes a change in sign of the controller output. This means, a negative controller output causes the addition of a positive value for the set point of the cascaded parameter and vice versa. The pH regulation with base and CO2 instead of acid is a classic example of this: to reduce the pH, the CO2 flow rate (CO2Flow parameter) needs to increase. The fact that the Negative function has been switched on is illustrated by the triangle symbol on the arrow that indicates the direction of the cascade control. This arrow shape can be seen both in the Cascade menu as well as in the Controller menu. 9.8.4 Special Configurations For bioreactors with gassing strategy “High End” (configuration with several mass flow controllers for flow control and gas mix) the gases to be used e.g. Air Flow, N2 Flow and O2 Flow, must be assigned to both parameters that control the gas mixture, i.e. parameters GasMix and GM Flow, in the cascade configuration. For this purpose, setup the following cascades additionally to the desired cascade configuration, if the appropriate parameters are present:  Parameter Air Flow as slave controller to parameter GM Flow  Parameter O2 Flow as slave controller to parameter GasMix  Parameter N2 Flow as slave controller parameter GasMix Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 208 of 257 11 April 2019 If parameters O2 Flow and N2 Flow are present, then they are setup as a parallel cascade below parameter GasMix. To make a distinction between the allocation of these parameters and regular cascade elements, the connections are shown without arrow. 9.9 Pumps The pumps are controlled in accordance with the corresponding parameters: Standard  Acid pump (digital): in accordance with the pH parameter  Base pump (digital): in accordance with the pH parameter  Antifoam pump (digital): in accordance with the Antifoam pa- rameter  Feed pump (analogue): in accordance with the Feed parame- ter Optional  Feed 2 pump (analogue): in accordance with the Feed 2 pa- rameter Digital pumps have a set speed and are time controlled. I.e., they always run at the same speed in start/stop mode. Analogue pumps have a variable speed (0 % to 100 %). Both digital and analogue pumps are controlled within a range of 0 % to 100 %. Example  Analogue: 50 % of the maximum feed rate = pump runs at half speed.  Digital: 50 % of the maximum feed rate = pump runs during half the time Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 209 of 257 The following pump settings are possible:  Setting the pump speed for feed pump(s) and dosing/waiting time for the antifoam pump  Calibrating the pumps  Resetting the delivery rate manually to zero  Filling or emptying the pump hoses automatically INFORMATION This function is only present for the four standard pumps. For details on how to change the settings for the feed pump(s) and the antifoam pump refer to the appropriate chapters in chapter “Parameters”. Calibration, pump counter and automatic filling and emptying is described in detail in the following chapters. 9.9.1 Calibrating a Pump Calibrating a pump makes it possible to display and record the actual delivered volume. The delivery rate is indicated in millilitres. Not the following points:  Always use hoses of the same kind with the same dimensions for calibration and pumping media.  Pump calibration must be executed before sterilisation. Aid  Graduated measuring cylinder/jug or scale/balance and an empty vessel  Reagent bottle equipped with silicone hose, filled with the reagent to be delivered, the feed or a liquid which has the same viscosity INFORMATION To obtain precise results, the reagent bottle should be put on a scale which is linked to the bioreactor or to the bioprocess platform software eve® installed on a PC or laptop. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 210 of 257 11 April 2019 In order to calibrate a pump, e.g. the acid pump, proceed as fol- lows: Procedure 1. Connect the reagent bottle to the pump. 2. Place the output end of the hose in a measuring cylinder/jug. Or: Place the reagent bottle on a scale and tare to zero, place the output end of the hose in an empty vessel 3. Completely fill the hose using the rocker switch of the base pump. 4. Call up the main menu Batch and press Calibrate Acid. The Calibrate Acid Pump dialogue box appears with:  Line 1, instruction on: Insert/prepare tube.  Line 2, instruction: Push button to fill tube.  Line 3, instruction: Tare balance (or empty measuring cup).  Line 4, instruction: Select pump speed. 25%, 50%, 75% and 100% and input field %: to select the pump speed in percent or enter another value in %. After selecting one of the buttons, the Other button appears instead of the % input field. Selection can be cancelled using this button.  Line 5, instruction: Select calibration time. 30m, 1h, 6h, 12h and input field min: to select the calibration duration or enter another calibration duration in minutes.  OK: to confirm entries and start calibration.  Cancel: to cancel procedure and close the dialogue box without changes. 5. Select the desired pump speed in % or enter it via numeric keypad. INFORMATION To obtain most accurate results, the pump should be calibrated at the same speed as it is to be expected to run during fermen- tation/cultivation. 6. Select calibration duration or enter it manually via numeric keypad. 7. Press OK. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 211 of 257 Calibration is started. This is indicated by In progress… and time left…. in h/min/s shows elapsing time next to the now enabled Stop button.. Once the time has elapsed: the Calibrate Acid Pump Part 2 dialogue box appears with:  Enter Liquid Volume gr (or ml): to enter the delivered volume in mL or g.  Pump Factor: displays the calculated pump factor after entry of the delivered volume. The pump factor is always ≠ 1 with a calibrated pump.  OK: to confirm entries.  CANCEL: to cancel calibration and close the dialogue box without changes. 8. Enter the delivered liquid in mL or g via numeric keypad. The pump factor is calculated automatically and displayed in the Pump Factor view box. 9. Press OK. The dialogue box disappears; the calibration value is saved. Done at with date and time next to the Stop button indicates that and when the pump was calibrated. 9.9.2 Resetting the Pump Counter to Zero The number of pump revolutions and the delivered quantity (in mL) of calibrated pumps are displayed constantly during a running fermentation/cultivation process. The display remains in place after completion of the process (when the bioreactor is stopped) until a new fermentation / cultivation process is started again (when the bioreactor is started). The counter can also be reset to zero manually, proceed as fol- lows: Procedure 1. Call up the main menu Main. 2. Press the desired pump button, e.g. Acid. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 212 of 257 11 April 2019 The Acid pump properties dialogue box appears with:  Pump factor: displays the pump factor or allows to be changed manually. The pump factor is always ≠ 1 with a calibrated pump. INFORMATION Manual change of the pump factor leads to rejection of the previous calibration value.  Duration: displays number of pump revolutions  Value: displays delivered quantity in mL  Reset: to switch on the reset of the pump counter.  Cancel: to close the dialogue box without changes.  OK: to confirm entry and close the dialogue box. 3. Activate the counter reset via Reset. The function is switched on. 4. Press OK. The counter is reset to zero and the count starts from the beginning, as soon as the bioreactor is started. 9.9.3 Automatically Filling/Emptying Pump Hoses Fill/Empty Pumps in main menu Batch allows the automatic filling or emptying of the pump hoses of the four standard pumps when the bioreactor is in stopped state. For each pump, an individual filling/emptying duration can be defined. Note the following:  The pump duration of a pump should preferably be tested with the liquid which has the same or similar viscosity as the liquid to be pumped.  Observe hose lengths and hose sizes of the pumps and, if necessary, test the pump duration of each pump considering the same conditions mentioned above. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 213 of 257 Content of Fill/Empty Pumps Menu All pumps (All Acid Pumps etc.) listed on four lines, divided in two columns with:  Input fields Running time (sec): to enter the running duration of the pump in seconds, either for filling or for emptying.  Fill: to start automatic filling of the selected pump.  Empty: to start automatic emptying of the selected pump. Stop buttons are provided next to each of these buttons for immediately stopping the filling or emptying process.  OK: to close the menu. INFORMATION If a filling or emptying procedure is active, the remaining filling or emptying duration is displayed. The menu cannot be closed while at least one filling or emptying procedure is active. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 214 of 257 11 April 2019 9.10 SIP – Sterilisation in Place Sterilisation is always carried out according to the application and user specifications. General information about in situ sterilisation and possible methods are described in main chapter “Preparations before Fermentation/Cultivation”, chapter “In situ Sterilisation – General Infor- mation”. The following chapters describe the SIP processes (Sterilisation in Place) which are started on the touch screen software. WARNING The vessel is under pressure during sterilisation! Removing mounting parts or the vessel top plate lead to spurting out of liquids and/or rapid exhausting of gasses. This may cause severe chemical burns, burns or intoxication. Always ensure the vessel is unpressurised before manipulating on mounting parts or the vessel top plate. CAUTION Danger of scalding and burns due to contact with hot surfaces! The vessel, the pipework and their components can get hot during sterilisation and lead to burns! INFORMATION The following chapters describe full sterilisation process and the sterilisation process of the harvest/sample valve as built and configured on the pilot bioreactor Techfors-S with serial number S-129020. Sterilisation processes of the optional sample valve and the resterilisable feed line are not described here. 9.10.1 Full Sterilisation – Vessel Sterilisation The water in the vessel jacket is heated up by steam injection for full sterilisation of the vessel. The steam generated by the liquid in the vessel sterilises the filters for inlet air and exit gas at the same time. The harvest/sample valve (bottom valve) is sterilised separately, see chapter “SIP Harvest/Sample Valve - Sterilisation of the Harvest/Sample Valve”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 215 of 257 9.10.1.1 Process Sequences The following table shows the different process sequences in the left column and describes the dialogue boxes and status messages which will appear in the touch screen software in the right column. Sequence Description Configuration Dialogue box Full sterilisation: configuration for process configuration (see next chapter) User interaction required Dialogue box Full sterilisation: user interaction required with in- struction(s) Start delay Status: starting inhibitor + time left in h:min:s Sequence is skipped, if function is switched off. Heating up to degassing temperature Status: heating up to degas. temp. + setpoint temperature in °C Sequence is skipped, if value for degassing time = 0. Degassing at degassing temperature Status: degassing at temp. + setpoint temperature in °C + time left in h:min:s Sequence is skipped, if value for degassing time = 0. Heating up to sterilisation temperature Status: heating up to steril. temp. Sterilisation at sterilisation temperature Status: sterilising at temp. + setpoint temperature in °C + time left in h:min:s Cooling down to 103 °C Status: cooling down to 103 °C Cooling down to 80 °C Status: cooling down to 80 °C Immediate process abortion is possible at temperature < 80 °C. Cooling down +10 °C above inoculation temperature Status: cooling down to inoc. temp. +10 °C Holding phase Status: holding inoc. temp. + setpoint temperature in °C Holds inoculation temperature until fermentation/cultivation is started or full sterilisation is stopped. Process end Status: STOP activated at OR fermentation started at with date and time. Degassing All liquids contain some physically dissolved air at room temperature, which escapes when heating up. Liquid is free of gas at 121 °C. If the medium is quickly and continuously heated up to the selected sterilisation temperature, the escaping gas in the medium cannot escape fast enough, so that foam will strongly build up during sterilisation. Foam can be prevented or reduced by systematic degassing. Duration time and temperature at which the medium shall be degassed are defined in the configuration dialogue, see chapter “Process Configuration”. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 216 of 257 11 April 2019 General information about the process sequences All buttons for processes that cannot be simultaneously running are deactivated. Depending on the current phase of the process, values change and are displayed in the main menu Controller. The actual state of valves, actuators, in and outputs is displayed in the submenu Valves. INFORMATION Should the sterilisation temperature fall below the set point, the message temp low will appear. The countdown stops until the temperature has reached the set point again to proceed with the process. 9.10.1.2 Process Configuration Input field Value range Unit Stirrer speed 10 to 1000 min1 Sterilisation temperature 110 to 125 °C Sterilisation time 10 to 60 min Degassing temperature to 98 °C Degassing time 0 to 60 min Cooling flow Cooling flow during cooling phase to prevent vacuum in the vessel. 10.0 to 20.0 L/min Inoculation flow Air flow during inoculation phase (holding phase) 0 / 0.2 to 20.0 L/min Inoculation temperature up to 79 °C Delay Start (Process start delay) ON / OFF Hours 0 to 99 h Minutes 0 to 59 min (Starting process in … hours … minutes) 9.10.1.3 Process Start Before starting the process, check and ensure that:  All required services are available and activated.  All services have the correct connection pressure.  The mechanical seal is lubricated. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 217 of 257 ATTENTION A mechanical seal, which has not been adequately lubricated is destroyed when running dry.  Antifoam sensor is removed.  Inoculation needle(s) is/are removed.  Push valve(s) is/are closed. To start the process, proceed as follows: Procedure 1. Call up main menu Batch and press Full Sterilisation. The configuration dialogue box appears. INFORMATION The configuration dialogue contains more or less input fields depending on the equipment’s configuration. The figure to the left serves as an example. 2. Enter the desired values via numeric keypad and activate delayed start, where appropriate. Once all values are entered: 3. Press OK. 4. Enter the desired values via numeric keypad and activate delayed start, where appropriate The Full sterilisation: user interaction required dialogue box appears with the instruction, to execute the following step in the procedure: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 218 of 257 11 April 2019 a) Turn valve 805 in Position STER. Once executed: 5. Press OK. The program automatically runs through the different process sequences until the holding phase of the configured inoculation temperature. 9.10.1.4 Process End Inoculation temperature is held until the sterilisation is stopped via the Stop button or by starting the bioreactor (fermentation/cultivation) via the Start button. The process end is either indicated by STOP activated at or fermentation started at with date and time. 9.10.1.5 Process Abortion The process is aborted by pressing the Stop button next to the Full Sterilisation button. For safety reasons, the process can only be aborted immediately at a temperature < 98 °C. This means, if the temperature is ≥ 98 °C, the cooling sequence will start first and be indicated appropriately. Process abortion is indicated by aborted at with date and time. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 219 of 257 9.10.2 SIP Harvest/Sample Valve – Sterilisation of the Harvest/Sample Valve The harvest/sample valve can be sterilised as often as required with clean steam. This procedure is independent of the full sterilisation process. Approximately 10 to 20 minutes of sterilisation before and after sampling are usually sufficient. After sterilisation, the harvest/sample valve should cool down before sampling. The process is started and stopped in the touch screen software and the sterilisation duration is entered in the configuration dialogue. But steam inlet is manually regulated via valve 203, which is located on the steam hose line of the harvest/sample valve. INFORMATION All buttons for processes that cannot be simultaneously running are deactivated during the process. 9.10.2.1 Process Sequences and Process Configuration The following table shows the different process sequences in the left column and describes the dialogue boxes and status messages which will appear in the touch screen software in the right column. Sequence Description Configuration Dialogue box Sterilisation harvest/sample valve: configuration User interaction required Dialogue box Sterilisation harvest/sample valve: user interaction required with instructions Sterilisation Status: sterilisation + time left in h:min:s User interaction required Dialogue box Sterilisation harvest/sample valve: user interaction required with instruction End of sterilisation Status: finished after + time in h:min:s Duration of sterilisation is set in the configuration dialogue box: Input field Value range Unit Sterilisation time 10 to 60 min 9.10.2.2 Process Start and End Before starting the process, check and ensure that:  All required services are available and activated.  All services have the correct connection pressure. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 220 of 257 11 April 2019  Harvest/sample valve 201 is closed.  Steam hose is connected to valve 203. To start the process, proceed as follows: Procedure 1. Call up main menu Batch and press SIP Harvest/Sample Valve. The configuration dialogue box appears. 2. Enter the desired values via numeric keypad. 3. Press OK. The Sterilisation harvest/sample valve: user interaction required dialogue box appears with instructions, to execute the following steps in the procedure: a) Connect the steam trap to harvest/sample valve 201. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 221 of 257 b) Open valve 203 (Steam harvest/sample valve). Once executed: 4. Press OK. The sterilisation sequence (countdown) begins. Once sterilisation time has elapsed, the second Sterilisation harvest/sample valve: user interaction required dialogue box appears with the instruction to execute the following step in the procedure: a) Close valve 203 (Steam harvest/sample valve). Once executed: 5. Press OK. The process is finished. 9.10.2.3 Process Abortion The process can be aborted any time by pressing the Stop button next to the SIP Harvest/Sample Valve button. The same dialogue box like for the process end appears with the instruction for closing valve 203. That the process has been aborted and after how much time is indicated with aborted after + time in h/min/s. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 222 of 257 11 April 2019 9.11 Starting & Stopping the Bioreactor, Shutting Down the System 9.11.1 Starting the Bioreactor (Fermentation/Cultivation) Before you start the process, check and ensure that:  All required services are available and activated.  All services have the correct connection pressure.  The mechanical seal is lubricated. ATTENTION A mechanical seal, which has not been adequately lubricated is destroyed when running dry.  Inoculation needle(s) is/are mounted  Push valve(s) is open To start the bioreactor, proceed as follows: Procedure 1. Call up main menu Batch and press Start. The configuration dialogue box appears containing the controlled parameters. Setpoint settings of the previous fermentation/cultivation are visible here. INFORMATION The configuration dialogue box contains more or less process parameters depending on the equipment’s configuration. The figure below serves as an example. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 223 of 257 2. Change setpoint settings, if necessary and switch parameters on. INFORMATION The bioreactor is started with the settings in the configuration dialogue box. Changes to these settings are saved and transferred to the next configuration dialogue box. If setpoint values are changed or parameters are switched on /off whilst the bioreactor is running, these settings are only adopted for the current fermentation/cultivation process. Once all settings are made: 3. Press OK. The Bioreactor: user interaction required dialogue box appears with the instruction to execute the following step in the procedure: Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 224 of 257 11 April 2019 a) Turn valve 805 in position OP (= operation). Once executed: 4. Press OK. Once the bioreactor (fermentation/cultivation) is started, the status message in progress since indicates that and since when (days/hours/minutes/seconds) the bioreactor is running. All buttons for starting processes that cannot be simultaneously running are deactivated. CAUTION Danger of scalding and burns due to contact with hot surfaces! The vessel, the pipework and their components can get hot during fermentation/cultivation and lead to burns!  Current values and controller outputs are visible in the main menu Controller.  A recording of the current values and a diagram are visible in the main menu Trends. WARNING The vessel may be under pressure during operation! Removing mounting parts or the vessel top plate lead to spurting out of liquids and/or rapid exhausting of gasses. This may cause severe chemical burns, burns or intoxication. Always ensure the vessel is unpressurised before manipulating on mounting parts or on the vessel top plate. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software 11 April 2019 Page 225 of 257 9.11.2 Stopping the Bioreactor To stop the bioreactor, proceed as follows: Procedure 1. Call up main menu Batch and press Stop next to the Start but- ton. The Bioreactor operation: user interaction required dialogue box appears with:  Instructions Bioreactor operation will be stopped and Press OK to confirm.  OK: to stop the bioreactor and close the dialogue box.  Cancel: to close the dialogue box without changes. 2. Press OK. The bioreactor is stopped. Stopped after with display of d/h/min/s below Start indicates after how much running time the bioreactor was stopped. 9.11.3 Shutting Down the System, Switching Off the Equipment To shut down the system and to switch off the equipment, proceed as follows: Procedure 1. In main menu System, press Shutdown. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Operation Touch Screen Software Page 226 of 257 11 April 2019 Dialogue box Confirmation appears with:  Question: Do you want to shutdown the system?  OK: to shut down the system.  Cancel: to close the dialogue box without changes 2. Press OK. The system is shut down and switched off. ATTENTION Switching off at the main switch without previously stopping the bioreactor and/or shutting down the system on the operating panel may lead to damage of the operating panel! When the screen is dark: 3. Turn the main switch counter clockwise (quarter turn) in position 0/OFF. 4. Close/shut off supply lines and ensure they are pressure-free. WARNING The vessel may be still pressurised after switching off at the main switched due to stored energy! Check the vessel pressure on the manometer and, if required, put the vessel into non-pressurised condition before every manipulation on the vessel and its components Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance 11 April 2019 Page 227 of 257 10 Cleaning and Maintenance The following chapters describe in detail how the vessel, the top plate and accessories are cleaned and, as required, stored. Furthermore, the chapter contains a maintenance plan and corresponding descriptions for the procedures to be performed by the operator. 10.1 Cleaning Agent and Disinfectant Intended use Allowed products/tools For vessel with slight con- tamination Water Cleaning agent for denaturation of proteins 0.1 N NaOH Cleaning agent for smaller component parts (e.g. exit gas cooler) Ultrasonic bath Cleaning agent for sur- faces Water Disinfectant for surfaces Ethanol, 70 % 10.2 Cleaning the Vessel After ending fermentation/cultivation followed by in situ sterilisation for decontamination (depending on user specifications), the vessel should be cleaned. Depending on the degree and kind of contamination, rinsing with water can be sufficient. If remains of foam or protein are adhering to the inside of the vessel, then the following procedure will ensure sufficient cleaning: Procedure 1. Carefully remove sensors from the ports and put aside in order to clean them separately according to the manufacturer’s specifications. 2. Fill the vessel with 0.1 N NaOH. 3. Close all vessel ports and fit the vessel top plate. 4. Start the bioreactor and stir strongly for 2 hours by using the stirrer function. Temperatures of e.g. 40 up to 60 °C improve the cleaning action, prolong stirring duration as necessary. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance Page 228 of 257 11 April 2019 5. Stop the bioreactor, shut down the system and switch off the equipment. 6. Empty the vessel. 7. Thoroughly rinse the vessel with water. Repeat the procedure, if necessary. If the vessel is not used again for the next fermentation right after, sufficient air circulation in the vessel should be ensured. 10.3 Cleaning the Vessel Top Plate Depending on the application, separate cleaning of the vessel top plate may be necessary. To clean the vessel top plate thoroughly, proceed as follows: Procedure 1. Lift up and remove the vessel top plate and put it onto an appropriate storage area. INFORMATION A detailed description how to remove the vessel top plate can be found in chapter “Removing the Vessel Top Plate” of the main chapter “Preparations before Fermentation/Cultivation”. Observe the safety advice and follow the instructions stated in there. 2. Dismantle component parts. 3. Carefully rinse all component parts with water. Use 0.1 N caustic soda solution as necessary. 4. Carefully rinse the vessel to plate with water or wipe it with a damp cloth or sponge. Use 0.1 N caustic soda solution as necessary. 5. Check vessel top plate sealing (O-ring) and O-rings of mounting parts on damages and replace them as necessary. 6. Leave the vessel top plate and the mounting parts to dry or dry wipe them. 7. Re-assemble clean and dry mounting parts into the also clean and dry vessel top plate. 8. Store the vessel top plate clean and dry in a location where it is secure and unable to fall or be damaged in passing, if not used right after for next fermentation/cultivation. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance 11 April 2019 Page 229 of 257 10.4 Dismantling and Cleaning the Exit Gas Cooler To remove the exit gas cooler from the vessel top plate in order to dismantle and clean it, proceed as follows: Procedure 1. Remove the red pressure hoses from the exit gas cooler (the red pressure hoses are not visible on the picture to the left). INFORMATION The pressure hoses for water inlet/outlet of the exit gas cooler are secured with hose clamps. 2. Remove the two clamps before and after the exit gas filter along with the exit gas filter itself. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance Page 230 of 257 11 April 2019 Ensure that the flat gaskets between the flanges do not get lost. 3. Remove the clamp between the bottom flange of the exit gas cooler and the flange on the vessel top plate 4. Remove the exit gas cooler from the vessel top plate and put it aside on a work space, where it cannot fall off. Ensure the flat gasket does not get lost. 5. Carefully lift the inner plates out of the main body of the exit gas cooler. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance 11 April 2019 Page 231 of 257 6. Put the disassembled exit gas cooler (inner plates and main body) into 0.1 N NaOH for 4 hours. 7. Thoroughly rinse the exit gas cooler with water. 8. Put the exit gas cooler into an ultrasonic bath for 2 to 5 minutes. 9. Wash the exit gas cooler through with 70 % Ethanol. 10. Thoroughly rinse the exit gas cooler with distilled water. 11. Let it dry in a clean place and re-assemble when dry. 10.5 Cleaning Reagent Bottles, Hoses and Component Parts Reagent bottles and hoses are separately autoclaved before cleaning. For details refer to main chapter “Fermentation/Cultivation”, chapter “Sterilisation after Fermentation/Cultivation” After autoclaving and cooling down, proceed as follows: Procedure 1. If appropriate, carefully empty the reagent bottles, and dispose of the liquid respecting the internal safety regulations. 2. Thoroughly rinse reagent bottles, hoses and component parts such as e.g. inoculation needles, push valves etc. with water. 3. Check silicone and pump hoses on damage and replace as necessary. INFORMATION Depending on user specifications, the hoses are replaced after every use. 4. Check reagent bottles and its components on damage and replace as necessary. 5. Check O-rings on component parts and lid seals on the reagent bottles on damage and replace as necessary. 6. Leave to dry the reagent bottles, hoses and component parts on a clean surface. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance Page 232 of 257 11 April 2019 10.6 Cleaning the Sensors Cleaning and storage of the sensors is described in the separate documentation of the corresponding sensor manufacturer. Strictly follow these instructions. The antifoam and/or level sensor from the equipment manufacturer is cleaned and maintained like any other component part (inoculation needles, push valves etc.). If not used for next fermentation/cultivation, it is stored when clean and dry. 10.7 Cleaning Surfaces of the Instrumentation Cabinet and Operating Unit If required, the surfaces of the instrumentation cabinet and the operating unit can be cleaned. ATTENTION Take the protection classes IP43 (instrumentation cabinet) and IP66 (operating unit) into account when cleaning! Proceed as follows: Procedure 1. Ensure the equipment is switched off at the main switch, switch off, where applicable. 2. Disconnect from mains supply. 3. Wipe the surface of the instrumentation cabinet and of the operating unit with a soft, damp cloth. Use an appropriate (non-aggressive!) disinfectant as neces- sary. 4. Clean the screen of the operating unit with a wipe suitable for computer or laptop screens. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance 11 April 2019 Page 233 of 257 10.8 Maintenance Plan WARNING Non-compliance of this maintenance plan contains a high risk! It is the responsibility of the user, that this maintenance plan is complied with. Non-compliance will lead to exclusion of liability (see General Terms and Conditions). The required maintenance for reliable operation is described in the following chapters. Reduce the maintenance intervals in case increased abrasion is detected during regular checks. Contact the manufacturer for questions concerning maintenance. For contact details, see page 2. To be carried out by operator Interval Maintenance work Before every operation Check hose lines and connections. Check all O-rings and rubber seals and replace if necessary. Check the reagent bottles and any other work tool made of glass on intactness; replace if necessary. Lubricate the mechanical seal. Check the filters with a filter test device, if available. After every fermentation/culti- vation Sterilise and clean the vessel, the vessel top plate, built in parts of the vessel top plate, reagent hoses and bottles. Proactively replace silicone hoses (frequency depends on application). After 20 – 50 sterilisations (rough recommendation) Replace air filters. Reduce or enhance maintenance interval as necessary. Every 6 months Replace all pump and silicone hoses on the reagent bottles. As required Clean the surfaces of the instrumentation cabinet and the operating unit. To be carried out by qualified personnel Interval Maintenance work Every 6 months Check and calibrate measuring sections (temperature, pH, etc.) with a sim- ulator. Replace all O-rings and rubber seals, reduce maintenance interval as nec- essary. As required Replace hoses and hose connections. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance Page 234 of 257 11 April 2019 To be carried out by qualified personnel from the manufacturer Interval Maintenance work When signs of wear or damage are apparent or at maintenance intervals defined by the operator of the equipment. Replace mechanical seal. INFORMATION For details about maintenance of parts and peripheral devices not made by the equipment manufacturer refer to the separate documentation from the manufacturer of these components. Read these and strictly follow instructions stated in there. 10.9 Lubricating the Mechanical Seal The silicone hose on the drive hub must always be filled with liquid (glycerine!) to ensure the mechanical seal is lubricated. ATTENTION Risk of loss of property due to the mechanical seal running dry! A mechanical seal, which has not been adequately lubricated is destroyed when running dry. To lubricate the mechanical seal proceed as follows: Procedure 1. Pull off the longer piece of hose from the coupling on the shorter piece. 2. Fill a syringe with glycerine 3. Plug the syringe onto the open hose end. 4. Fill glycerine into the hose. 5. Plug the longer piece of hose onto the coupling of the shorter piece. If glycerine has come off the hose, wipe off as necessary. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Cleaning and Maintenance 11 April 2019 Page 235 of 257 The following points need to be observed  A small loss of glycerine is normal during operation of the vessel. For this reason, it is recommended to top up with glycerine regularly, e.g. after several fermentations or every time the top plate is removed for cleaning. Provided the mechanical seal is in normal operational order, after topping up of a few cubic centimetres of glycerine will be visible in the hose where the glycerine is added.  Due to normal wear of the mechanical seal – which is made of hard carbon and silicon carbide – the glycerine will show a dark discoloration which is visible in the silicone hose. This discoloration is perfectly normal and should not be understood as a sign of progressive wear of the mechanical seal.  If glycerine loss is significant during or between fermentations, this might indicate a misaligned or defective mechanical seal. A significant glycerine loss can be detected if much more glycerine than usual is necessary for topping up the chamber and/or sudden dark discoloration of the culture liquid, due to glycerine running down the stirrer shaft.  If a significant loss of glycerine is observed, the manufacturer must be contacted to have the seal checked by qualified personnel from the manufacturer or local dealer to re-align or replace the seal.  When replacing the top plate after cleaning, leave it for some time before checking the level of the glycerine in the mechanical seal. This allows for any possibility that tilting of the top plate has temporarily drained some glycerine from its correct location. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences Page 236 of 257 11 April 2019 11 Interferences The following section describes possible reasons for interferences and how to resolve them. Reduce the service intervals in correspondence with the actual loads if interferences become increasingly common. Contact the manufacturer for interferences that cannot be resolved by following the above instructions. For service contact details, see page 2. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences 11 April 2019 Page 237 of 257 11.1 Interferences Basic Operation and Operating Unit Interference Equipment does not work, power display POWER is not illuminated, no display. Possible Causes Remedy By Power cable not connected. Connect power cable. Operator Main switch switched off. Switch main switch on. Operator Fuses blown Check fuses of mains connection. Check fuses of basic unit. Qualified elec- trician Break in power connections. Check power plug. Check power cable on damages or kinks. Check socket and cable on basic unit. Qualified elec- trician Interference Power display POWER is illuminated, no display. Possible Causes Trouble shooting By Monitor of operating unit is switched off at the ON/OFF key. Switch the monitor of the operating unit on at the ON/OFF key. Operator Power supply cable not connected to the operating unit. Plug the power supply cable in at the DC socket on the rear side of the operating unit Operator Power supply cable not connected to controller inside the instrumentation cabinet. Connect the power supply cable to the controller inside the instrumentation cabinet. Qualified elec- trician Interference No communication between equipment and operating unit. Alarm no communication. Possible Causes Remedy By iDDC-bus cable (display cable) not connected to the operating unit. Plug the round connector of the iDDC-bus cable in at the COM1 socket (labelled with RS-485) on the rear side of the operating unit. Operator iDDC-bus cable (display cable) is not connected to the controller inside the instrumentation cabinet. Connect the iDDC-bus cable (display cable) to the controller inside the instrumentation cabinet. Qualified elec- trician Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences Page 238 of 257 11 April 2019 11.2 Interferences Drive System Interference Motor does not start Possible Causes Remedy By Parameter Stirrer is not switched on. Switch parameter Stirrer on. Operator Stirrer setpoint = 0. Set Stirrer setpoint > 0 Check value in Deadband of parameter option PID: Value must be = 0 (zero) Operator Parameter pO2 is switched on and set to work with stirrer. (Option Cascade in parameter pO2) Switch Cascade off and test operation via parameter Stirrer. Operator Interference Motor does not start, parameter Stirrer is turned on, Cascade in pO2 is not activated. Possible Causes Remedy By Motor cable not properly connected Check motor cable connection, connect properly as necessary. Qualified per- sonnel Motor is/was overheated or mains voltage of the motor is too low. Turn the equipment off at the main switch. Wait for approx. 20 seconds. Turn the equipment on at the main switch. If this does not resolve the problem, see next sec- tion. Operator Turn the stirrer drive (parameter Stirrer) off at the operating panel. Open the instrumentation cabinet. Check the LED display of the motor controller:  Error Code M: motor overheated  Error Code Z: low mains voltage. Press the Reset button of the frequency converter. Close the instrumentation cabinet. Turn on parameter Stirrer. Qualified elec- trician Interference Motor control is erratic Possible Causes Remedy By Wrong settings in PID option of parameter Stirrer Reset default settings in PID menu. Operator Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences 11 April 2019 Page 239 of 257 11.3 Interferences Temperature Control System Interference No temperature control. Possible Causes Remedy By Parameter Temp (temperature) is not switched on Switch parameter Temp (temperature) on Operator Interference No cooling or inadequate cooling. Possible Causes Remedy By No water supply or inadequate flow. Ensure water supply is turned on and flow is suffi- cient. Operator Incorrect Neg. factor (negative factor) in option PID of parameter Temp (temperature) Check Neg. factor (negative factor) in option PID of parameter Temp (temperature): Value must be posi- tive Operator Interference Temperature drifts up or down over time Possible Causes Remedy By Incorrect settings in option PID of parameter Temp (temperature) Check settings in option PID of parameter Temp (temperature) and adjust as necessary (especially P-term). Operator Interference Alarm no water detected in temperature control system. Temperature control is switched off, circulation pump and heating are deactivated, and active process continues running. Possible Causes Remedy By Temperature control circuit is not filled Fill the temperature control circuit Qualified per- sonnel Interference Negative temperature value Possible Causes Remedy By Broken cable or faulty sensor Replace temperature sensor Infors service technician or licensed dealer Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences Page 240 of 257 11 April 2019 11.4 Interferences pH System Interference No display or incorrect display of pH Possible Causes Remedy By Sensor cable is twisted or kinked or not properly connected. Check and ensure that the sensor cable is not kinked or twisted. Connect the sensor cable properly as necessary. Operator pH sensor is not calibrated Test calibration with pH4 and pH7 buffers Operator Temperature compensation function is switched off. Switch temperature compensation function on in normal use. Operator pH drift during long-term fermenta- tion Recalibrate pH with offline values (“pH Sensor Re- calibration”). Operator Faulty pH sensor Read documentation from sensor manufacturer Operator Interference No pH control Possible Causes Remedy By Parameter pH is not switched on. Switch parameter pH on. Operator No control in Deadband in option PID Check Deadband in option PID of parameter pH: switch off or enter a small value. Operator No corrective reagents. Check and refill bottles, if necessary Check connection with vessel. Connect properly, if necessary. Ensure that reagent hose line(s) is/are not clamped off. Remove clamp(s), if necessary. If working with push valve(s): Check and ensure that the push valve(s) is/are open, open as necessary. Operator Acid or/and Base pump(s) does/do not operate properly. Check operation of the pump using the rocker switch. Check pump hose type and replace if necessary. Operator Interference pH value drifts up and down over time or acid and base are added almost continuously in turn. Possible Causes Remedy By Incorrect settings in option PID Check settings in option PID of parameter pH and adjust as necessary (especially P-term). Operator Incorrect strength of reagents. Ensure that reagents are of roughly equal strengths in terms of effect (i.e. 0.1 to 2.0 molar). Operator Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences 11 April 2019 Page 241 of 257 11.5 Interferences pO2 System (Dissolved Oxygen) Interference No display or incorrect display of pO2 Possible Causes Remedy By Sensor cable is twisted or kinked or not properly con- nected. Check and ensure that the sensor cable is not kinked or twisted. Connect the sensor cable properly as necessary. Operator pO2 sensor is not polarised. Polarise the pO2 sensor Operator Faulty sensor Check the calibration of the pO2 sensor. Consult the documentation of the sensor manufacturer. Operator Interference No pO2 control Possible Causes Remedy By Parameter pO2 is not switched on. Switch parameter pO2 on. Operator Incorrect settings in option Cascade of parameter pO2 Check set up of cascade, adjust settings, if necessary. Operator No gas flow into vessel. Check supply connections. Check and ensure that gas supply is turned on. Turn on, if necessary. If present, check and ensure that the rotameter valve is fully open. If present, ensure the Flow parameter(s) is/are switched on and flow rate is not set too low a value, adjust as neces- sary. Operator Interference pO2 control is unstable Possible Causes Remedy By Incorrect settings in option PID of parameter pO2 Check settings and adjust as necessary, especially P-term and Deadband. Deadband must be 0. Operator Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences Page 242 of 257 11 April 2019 11.6 Interferences Antifoam Control Interference Foam is not sensed Possible Causes Remedy By Sensor cables are not properly con- nected Connect properly as necessary. Operator Interference Foam is always/frequently detected Possible Causes Remedy By Sensor cables are not properly con- nected Connect properly as necessary. Operator Transparent insulation of antifoam sensor damaged Replace transparent insulation of antifoam sensor Qualified personnel from the manufac- turer Interference Antifoam pump does not work Possible Causes Remedy By Parameter Antifoam is not switched on. Switch the parameter on. Operator Dosing time of parameter Antifoam = 0 (zero). Set dosing time of parameter > 0. Operator Interference No or inadequate reagent. Possible Causes Remedy By Reagent bottle empty Refill bottle, if necessary. Operator Wrong antifoam agent or wrong con- centration Replace agent, if necessary. Operator Hose line blocked or clamped off. Check hose line connection between reagent bottle and vessel, connect properly as necessary. Open/remove hose clamp, if necessary. Operator If working with push valve: valve closed Open the push valve as necessary. Operator Antifoam pump does not operate properly Check operation using the rocker switch. Check hose type and replace if necessary. Operator Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences 11 April 2019 Page 243 of 257 11.7 Interferences Feed and Pump Interference No feed inlet Possible Causes Remedy By Feed parameter is/are not switched on. Switch Feed parameter(s) on. Operator Setpoint of Feed parameter(s) = 0 (zero). Enter setpoint of Feed parameter(s) > 0 Operator Reagent bottle empty Check and refill bottle, if necessary. Operator Feed hose line blocked Check connection with vessel. Connect properly if neces- sary. Check and ensure that hose line is not clamped off. Remove clamp, if necessary. Operator If working with push valve: valve is closed Open the push valve Operator Feed pump does not operate properly Check pump operation using the rocker switch. Check hose type and replace if necessary. Operator Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences Page 244 of 257 11 April 2019 11.8 Interferences Gassing System Interferences No gassing Possible cause Remedy By The in-house gas supply has been interrupted. Stop the bioreactor (fermentation/cultivation). Check the in-house gas supply and switch it on, if necessary. Operator Gasmix and/or Flow parameter(s) is/are not activated. Setpoint(s) in the Gasmix and/or Flow parameter(s) = 0 Activate the Gasmix and/or Flow parameter(s) as necessary. Set the setpoint(s) of the Gasmix and/or Flow parameter(s) > 0 as necessary. Operator Inlet air or exit gas escapes via twist valves on the filters. Check whether the twist valves are closed, close as necessary. Operator Inlet air filter blocked. Replace the inlet air filter under sterile conditions. Operator Interference The desired gas flow rate is not reached. Possible cause Remedy By Inlet air or exit gas filter blocked. Replace the filter under sterile conditions. Operator Wrong connection pressure of gas(es). Check connection pressure of gas, adjust as neces- sary. Operator Interference Sudden increase in evaporation losses in the culture vessel. Possible cause Remedy By The exit gas cooler does not cool. Check the water supply to the exit gas cooler, restore it if necessary. Check and ensure that the Cooler valve (118) in main menu Main is open (valve symbol is green) and set to automatic (Auto) Operator Techfors-S – Vessel Volume 15 L – Serial No. 129020 Interferences 11 April 2019 Page 245 of 257 11.9 Returning for Repair The provider must return the equipment or the faulty component part(s) to the manufacturer if, after consulting the service department of the local dealer or the manufacturer, on-site diagnosis and/or repair is not possible. INFORMATION When returning the equipment, the component part or accessory for repair, it is required for the safety of all parties involved and because of legal provisions that a lawful declaration of decontamination is present. Refer to main chapter “Safety and Responsibility”, chapter “Declaration of Contamination” for details. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Disassembly and Disposal Page 246 of 257 11 April 2019 12 Disassembly and Disposal The equipment must be disassembled and disposed of in an environmentally friendly manner if it is no longer in use. INFORMATION When returning the equipment for 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 present. Refer to main chapter “Safety and Responsibility”, chapter “Declaration of Contamination” for details. 12.1 Disassembly Prior to disassembly:  Switch off the equipment and lock any isolation switch in the ‘off’ position.  Physically disconnect the main energy supply from the equipment and wait for components to fully discharge.  Remove and dispose of all additional consumable items, auxiliary components and/or spent processing material in an environmentally acceptable manner. Clean and disassemble component parts professionally with regard to any local regulations concerning employment and environmental protection. If possible, separate materials. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Disassembly and Disposal 11 April 2019 Page 247 of 257 12.2 Disposal Recycle disassembled components if no agreement is made concerning reclaim or disposal.  Send metals for scrap.  Send plastic components for recycling.  Sort and dispose of the remaining components according their material composition. WARNING Electronic waste, electronic components, lubricants or other auxiliary materials/supplies are subject to hazardous waste regulations and may only be disposed of by registered specialist disposal firms. For disposal, the system units are to be disassembled and dismantled into individual material groups. These materials are to be disposed of according to the applicable national and local legislation. Local authorities or specialist disposal firms can provide information regarding environmentally acceptable disposal. If no special arrangements have been made for return, INFORS HT units with the required declaration of decontamination can be sent back to the manufacturer for disposal. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data Page 248 of 257 11 April 2019 13 Technical Data 13.1 Dimensions, Connections & Connection Values 1 Cooling water in (male adapter DN10) / 2.0 ± 0.5 bar 2 Cooling water out (male adapter DN10) / pres- sureless 3 Air out (male adaptor DN10) / pressureless 4 Air in (male adapter DN06) / ± 2.5 bar 5 O2 in (male adapter DN06) / ± 2.5 bar 6 N2 in (male adapter DN06) / ± 2.5 bar 7 Contaminated condensate out (male adapter DN09) / pressureless 8 Water in steam generator (male adapter DN09) / min. 3.0 bar (Water quality: CaCO3 concentration 0 mmol L-1 to max. 0.53497mmol L-1 ) 9 Total emptying steam generator (male adapter DN09) / pressureless Dimensions in mm Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data 11 April 2019 Page 249 of 257 13.2 Electrical Power Supply WARNING High leakage current! Earth connection essential before connecting supply! Bioreactor Description Value Unit Voltage 1 phase L1 + N (neutral) + PE (earth) 230 (± 5 %) V Frequency range 50 Hz Max. current 16 A Leakage current > 3.5 mA Steam Generator Description Value Unit Voltage 3 phases L1 + L2 + L3 + N (neutral) + PE (earth) 400 (± 5 %) V Frequency range 50 Hz Max. current 16 A INFORMATION The power supply of both, the bioreactor and the steam generator, must be individually made safe by the use of an FI-switch (or RCD – Residual Current Device) of the kind RCCB, Type B on the installation site. Detailed information about connection conditions as well as technical data, operation and maintenance of the steam generator can be found in the separate documentation from its manufacturer. Read the manual before initial operation of the equipment and strictly follow instructions stated in there. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data Page 250 of 257 11 April 2019 13.3 Specifications 13.3.1 Instrumentation Cabinet Description Value Dimensions W = 320 mm D = 450 mm H = 550 mm Protection class IP43 Material 1.4301 Peristaltic pumps Acid, Base, Antifoam, digital Feed, analogue 13.3.2 Operating Unit Description Value HMI Colour touch screen 12" Protection IP 66 13.3.3 Vessel Description Value Volume total Vessel: 30 L Vessel jacket: 2.0 L Working volume vessel Max. 20 L Min. 6.1 L Dimensions Height: 643 mm Inner diameter: 250 mm Ratio H/D: 2.5 : 1 Pressure range Vessel inside: -1 up to +3 bar Vessel jacket: 0 up to +3 bar Temperature range Vessel inside: -10 up to +150 °C Vessel jacket: -10 up to +150 °C Material in contact with media stainless steel 1.4404 or 1.4435 = AISI 316L with finish Ra ≤0.6 µm, electropolished Material not in contact with media stainless steel 1.4301 = AISI 304 with finish Ra ≤1.0 µm, electropolished Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data 11 April 2019 Page 251 of 257 Vessel top plate assem- bly  8 x 19 mm ports with Rd28x1/8"  2 Tri-Clamp DN10 ports, 1 for air inlet incl. diaphragm valve 805, 1 for exit gas cooler  1 x 10 mm port for temperature sensor (PT100)  1 connection flange for motor  4 recessed holes D = 8 mm for the baffles  2 x 4 mm holes for negative lead of the antifoam sensor  2 lifting handles  6 hexagon capped nuts and metal washers for fixing the vessel top plate Vessel upper section  2 INGOLD standard ports iD = 25 mm / G1-1/4", angled (3°)  2 Tri-Clamp 1-1/2” ports for CIP lances Vessel flange 6 threaded bolts M10 for fixing the vessel top plate Vessel jacket and INGOLD ports  1 INGOLD standard port iD = 25 mm / G1-1/4", horizontal  3 INGOLD standard ports iD = 25 mm / G1-1/4", angled (15°)  2 connections for the water inand outlet Vessel bottom 1 threaded nozzle for the harvest / sample valve Accessories  1 sight glass (115 x 15 mm)  1 vessel identification plate  4 baffles, removable, width 25 mm  1 ring sparger Harvest- /sample valve (bottom valve) Type NW25.5 incl. steam connection (ISO G1/8ʺ) Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data Page 252 of 257 11 April 2019 13.3.4 Temperature Description Value Heating / Cooling Steam heating provided by integrated steam generator 1) Tap water cooling (on site) Sterilisation Automatic with steam provided by integrated steam generator Temperature range Sterilisation 110 °C to 125 °C Fermentation 2) 20 °C to 79 °C Sensor Pt-100 class B, 1/3 DIN Accuracy temperature measuring & control (Fermentation) ± 0.3 °C at 10 to 60 °C ± 0.5 °C at > 60 °C 1) Detailed information about technical specifications, operation and maintenance of the steam generator can be found in the separate documentation from its manufacturer. 2) Min. fermentation temperature depends on ambient temperature, used cooling system, stirrer rotation speed and medium viscosity. 13.3.5 Stirrer Description Value Drive Top drive, with single mechanical seal Direction of rotation of drive shaft Clockwise (top view) Motor type AC servomotor, brushless Range of rotation speed 1) 10 to 1500 min-1 Accuracy control ± 5 min-1 at 10 to 1000 min-1 1 % setpoint at > 1000 min-1 1) Valid for liquid with viscosity similar to water, without gassing, with 2 Rushton impellers. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data 11 April 2019 Page 253 of 257 Impellers Description Value Number /Type 3 impellers, type Rushton, 6 blades, adjustable in height Dimensions A = 66 mm B = 13 mm C = 16 mm Material 316L, electropolished, Ra 0.8 µm 13.3.6 Gassing Description Value Gas(es) and entry Air, O2, N2 Gasmix entry via ring sparger or head room (deviation by means of valve 805) Solenoid valves Control Gas flow: 1 MFC Gasmix: 3 solenoid valves Mass Flow Controller (MFC) FIC801 Thermal mass flow controller Manufacturer: Vögtlin Instruments Type: red-y smart series Range: 0.2 – 20 L/min Accuracy: ± 3 % FS Connection thread: 1/4" Filter Manufacturer: Pall Type: Novasip, steam sterilisable Model: C3PFRP1A Max. pressure: 6.5 bar Max. temperature: +142 °C Retention rate: 0.2 µm Steam trap Type: balanced pressure thermostatic steam trap Material: stainless steel A detailed documentation about the filter body & element can be found in the separate documentation of the filter’s manufacturer PALL. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data Page 254 of 257 11 April 2019 13.3.7 Exit Gas Description Value Outlet Via exit gas cooler and exit gas filter into atmosphere Exit gas cooler Material: Stainless steel Filter Manufacturer: Pall Type: Novasip, steam sterilisable Model: C3PFRP1A Max. pressure: 6.5 bar Max. temperature: +142 °C Retention rate: 0.2 µm Steam trap Type: balanced pressure thermostatic steam trap Material: stainless steel Detailed information about the filter body & element can be found in the separate documentation from the manufacturer PALL. 13.3.8 pH Description Value Control Peristaltic pumps acid & base Control range pH 2 - 12 Measuring accuracy pH ± 0.1 Measuring system, analogue Type of sensor Conventional pH sensor potential measurement against reference Manufacturer METTLER TOLEDO Measuring range of sensor pH 2 - 12 For details about technical data, use and maintenance of the sensor see the separate documentation from the sensor manufacturer METTLER TOLEDO. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data 11 April 2019 Page 255 of 257 13.3.9 pO2 Description Value Control Cascaded stirrer Cascaded air flow Cascaded gasmix Cascaded O2 flow The functionality of the parameters depends on the hardware configuration of the equipment Control range 0 – 100 % Measuring accuracy 1 % FS Measuring system, analogue Type of sensor Traditional amperometric/polarographic pO2 sensor Manufacturer METTLER TOLEDO Measuring range sen- sor 0 % up to 150 % For details about technical data, use and maintenance of the sensor see the separate documentation from the sensor manufacturer METTLER TOLEDO. 13.3.10 Antifoam Description Value Sensor Conductive with dosing needle, adjustable in height Control Peristaltic pump (Antifoam) Range 0 / 100 % (OFF/ON) Sensor Description Value Length 350 mm Material  Parts in contact with medium: 316L  Surface finish of steel parts in contact with media: 0.8 µm, elec- tropolished O-rings EPDM Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data Page 256 of 257 11 April 2019 13.3.11 Pumps Description Value Type Peristaltic Digital 3 pieces Acid Base Antifoam Analogue 1 piece Feed Rotation speed Digital 150 min-1 / fixed rotation speed Analogue 150 min-1 / max. rotation speed, adjustable within range of 0 % up to 100 % Accuracy ± 1 min-1 13.3.12 Push Valve Description Value Process connection Rd28x1/8'' (19 mm port in vessel top plate) Parts in contact with medium Material: 316L, stainless steel Surface roughness: 0.8 µm, electro pol- ished O-ring EPDM 13.3.13 Pressure Indication Vessel Description Value Type Manometer Connection 19 mm port in vessel top plate Range 0 – 4.0 bar O-ring EPDM 13.3.14 Pressure: Safety Valves Description Value Safety valve vessel Triggering pressure: 3 bar g Manufacturer: Ramseyer Safety valve temperature control circuit Triggering pressure: 3 bar g Manufacturer: Ramseyer For details about technical data, use and maintenance of the safety valves refer to the separate documentation from the manufacturer Ramseyer. Techfors-S – Vessel Volume 15 L – Serial No. 129020 Technical Data 11 April 2019 Page 257 of 257 13.4 Operating Conditions Description Value Temperature range 5 °C – 35 °C Relative air humidity, non-con- densing 20 % - 90 % Min. distance from walls, ceilings 1) and other appliances 150 mm 1) The distance from the ceiling must be chosen in such a way that the vessel top plate including its built-in parts can easily be lifted from the vessel. 13.5 Operating Materials Application Permitted products Lubricant for the single mechanical seal (only bacterial system) Medicinal Glycerine 85 % Quality: PhEur 13.6 Emissions Description Value Units Noise emission <70 dB (A)