Manual Biostat Cplus Sbt6022-E

Operating Manual
BIOSTAT® Cplus
Fermentor / Bioreactor
85037-540-72 Vers. 02 | 2014

The enclosed CD contains the operating instructions
as a PDF file in various international languages.
If the CD is missing, you can obtain a copy form us

by specifying the order number:
Order number: 85037-543-23
Sartorius Stedim Biotech GmbH

Technical Editorial Department
August-Spindler-Strasse 11
37079 Goettingen, Germany
tech.pubs@sartorius-stedim.com
www.sartorius-stedim.com
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Content
Part A: BIOSTAT® Cplus

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
1.1 Presentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
1.2 Service Center . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2. Safety Instructions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
2.1 General Safety Instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
2.2 Informal Safety Measures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
2.3 Symbols Used on the Device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
2.4 Intended Use and Foreseeable Misuse . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
2.5 Residual Risks from Use of the Device . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.6 Danger Due to Electrical Power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
2.7 Danger Due to Components Under Pressure . . . . . . . . . . . . . . . . . . . . . . . 16
2.8 Hazards Arising from Bursting Culture Vessel . . . . . . . . . . . . . . . . . . . . . . 17
2.9 Danger Due to Gases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.9.1 Dangers Due to Oxygen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.9.2 Danger Due to Nitrogen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.9.3 Danger Due to Carbon Dioxide . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.10 Danger Due to Escaping Steam . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.11 Danger Due to Escaping Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.12 Danger Due to Hot Surfaces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.13 Danger Due to Rotating Parts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
2.14 Danger Due to Use of Incorrect Consumables . . . . . . . . . . . . . . . . . . . . . . 19
2.15 Personal Protective Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.16 Safety and Protective Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
2.16.1 Switch-Disconnector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
2.16.2 Safety Valves and Pressure Reducer . . . . . . . . . . . . . . . . . . . . . . . 21
2.17 Emergency Instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
2.18 Operator Responsibilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
2.19 Personnel Requirements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
2.20 Personnel Qualification Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
2.21 Personnel Responsibilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
2.22 Responsibilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
2.23 Unauthorized Personnel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
2.24 Training and Instruction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
3. Device Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
3.1 Overall views . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.2 Control | supply units . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
3.3 Aeration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.3.1 “O2 Enrichment” and “Gasflow Ratio” Aeration . . . . . . . . . . . . . 34
3.4 Pump Modules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
3.4.1 Performance Parameters and Features . . . . . . . . . . . . . . . . . . . . 36
3.4.2 External Pumps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
3.5 Culture Vessels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
3.5.1 Overview, 5-liter Culture Vessel . . . . . . . . . . . . . . . . . . . . . . . . . . 38
3.5.2 Overview, 10-liter to 30-liter Culture Vessel . . . . . . . . . . . . . . . . 40
3.6 Agitator and Agitation Drive . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
3.7 Steam Distributor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
3.8 Manual Pressure Control Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
4. Transportation and Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
4.1 Inspection Upon Acceptance by the Recipient . . . . . . . . . . . . . . . . . . . . . 44
4.1.1 Report and Document Transport Damage . . . . . . . . . . . . . . . . . . 44
4.1.2 Check Completeness of the Delivery . . . . . . . . . . . . . . . . . . . . . . 44
Contents 3
4.1.3 Packaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
4.1.4 Instructions for Transport within the Company . . . . . . . . . . . . . 45
4.2 Intermediate Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
5. Setup, Assembly, and Initial Commissioning. . . . . . . . . . . . . . . . . . . . . . . . . . . 47
5.1 Setup | Assembly. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
5.1.1 Installation Location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
5.1.2 Ambient Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
5.1.3 Acclimatization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
5.1.4 Workplace Requirements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
5.1.5 Supply Facilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
5.1.6 Disposal Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
5.1.7 Setup Examples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
5.2 Starting Up Temperature Control Circulation . . . . . . . . . . . . . . . . . . . . . . 53
5.3 Initial Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
6. Operation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
6.1 Safety Instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
6.2 Switching the Device On | Off . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
6.3 Triggering an Emergency Stop. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
6.4 Sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
6.5 Manual Pressure Control Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
6.6 Inserting Tubes into Peristaltic Pumps . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
6.7 Internal Spinfilter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
6.7.1 Design and Function . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
6.7.2 Installation and Connection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
6.8 Intake Units . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
6.8.1 Inoculation Port APC 19 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
6.8.2 Intake and Sampling Unit APC 25 . . . . . . . . . . . . . . . . . . . . . . . . 62
6.8.3 STT Quick Connector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
6.8.4 Inoculation Fittings and Septa . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
6.8.5 SACOVA Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
6.8.6 Correction Media Bottles. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
6.9 SVC 25 Sampling Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
6.9.1 Sampling Valve Standard, Design | Function. . . . . . . . . . . . . . . . 70
6.9.2 Containment Sampling, Design | Function . . . . . . . . . . . . . . . . . 72
6.10 Floor Drain Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
6.11 Dummy Plugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
6.12 Sterilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
6.12.1 Safety Instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
6.12.2 Splinter Protection Setup for 5-liter Culture Vessels . . . . . . . . . 77
6.12.3 Adjusting Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
6.12.4 Carry out Sterilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
6.12.5 Sterilizing the Double Mechanical Seal . . . . . . . . . . . . . . . . . . . . 78
6.13 Performing Processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
6.13.1 Sterile Test and Pressure-hold Test . . . . . . . . . . . . . . . . . . . . . . . . 83
6.13.2 Preparing the Bioreactor for the Process . . . . . . . . . . . . . . . . . . . 84
6.13.3 Inoculating the Culture Vessel . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
6.13.4 Completing the Process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
7. Cleaning and Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
7.1 Safety Instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
7.2 Assembly / Disassembly of the Motor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
7.3 Cover Lifting System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
7.3.1 Lifting the Cover Plate off of the Vessel . . . . . . . . . . . . . . . . . . . 88
7.3.2 Lowering the Cover Plate onto the Vessel . . . . . . . . . . . . . . . . . . 89
7.4 Disassembly / Assembly of the Cover Plate . . . . . . . . . . . . . . . . . . . . . . . . . 90
7.5 Installation of the Stirrer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
7.6 Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
7.6.1 Cleaning the Device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
4 Contents
7.6.2 Cleaning the Control Tower, Vessel, and Attachments . . . . . . . . 92
7.6.3 Intermediate Cleaning After Processes . . . . . . . . . . . . . . . . . . . . 93
7.6.4 Basic Cleaning and Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
7.7 Maintenance Instructions and Function Tests . . . . . . . . . . . . . . . . . . . . . . 93
7.7.1 Measures to Take after Maintenance . . . . . . . . . . . . . . . . . . . . . . 93
7.7.2 Device Maintenance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
7.7.3 Maintenance Intervals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
7.7.4 Seals. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
7.7.5 Replacing intake and exhaust air filters. . . . . . . . . . . . . . . . . . . . 98
7.7.6 Replacing the view glass lamp . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
7.7.7 Sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100
7.7.8 SACOVA valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104
7.7.9 Inoculation Fittings and Septa . . . . . . . . . . . . . . . . . . . . . . . . . . 105
7.7.10 Internal Spinfilter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106
7.7.11 STT Quick Connector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
7.8 Assembling and Adjusting the Sparger Pipe . . . . . . . . . . . . . . . . . . . . . . 108
7.9 Securing the Weigh Cells while Moving the Unit . . . . . . . . . . . . . . . . . . 108
8. Disruptions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
8.1 Safety Instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
8.2 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
8.2.1 Process-related Faults . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
8.2.2 Hardware-related Faults . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
8.2.3 Fault Table “Contamination” . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
8.2.4 Troubleshooting Table “Counter Cooling System” . . . . . . . . . . 112
8.2.5 Troubleshooting Table “Aeration and Ventilation” . . . . . . . . . . 112
9. Disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
9.1 General Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
9.2 Hazardous Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
9.3 Decontamination Declaration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
9.4 Decommissioning the Device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
9.5 Disposing of the Device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
10. Appendix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115
10.1 Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115
10.2 Connector pin assignment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
10.3 EC Declaration of Conformity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118
10.4 Dimensioning the Variable Area Flowmeters . . . . . . . . . . . . . . . . . . . . . . 120
10.5 Decontamination Declaration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
Part B: DCU4 System for BIOSTAT® Cplus

11. User Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
12. System Behavior During Start . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
13. Principles of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
13.1 “Main” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
13.1.1 Work Area . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
13.1.2 Header . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
13.1.3 Footer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
13.2 Display of Function Elements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
13.3 Overview of the Menu Buttons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
13.4 Overview of Selection Keys . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
13.5 Direct Function Keys for Selection of Submenus. . . . . . . . . . . . . . . . . . . 132
13.6 Selection Lists and Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134
14. “Main” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
14.1 General Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
14.2 Process Displays in the “Main” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
14.3 Direct Access to Submenus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
Contents 5
15. “Trend” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
15.1 “Trend” Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
15.2 Configuring the “Trend” Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138
15.2.1 Setting the Trend Display for Parameters . . . . . . . . . . . . . . . . . 138
15.2.2 Setting the Parameter Display Range . . . . . . . . . . . . . . . . . . . . 138
15.2.3 Resetting the Display Range . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
15.2.4 Setting the Trend Display Color: . . . . . . . . . . . . . . . . . . . . . . . . . 139
15.2.5 Defining a New Time Range as “Time Range” . . . . . . . . . . . . . . 139
16. “Calibration” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
16.2 pH Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
16.2.1 Calibration Sequence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
16.2.2 Recalibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 144
16.2.3 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145
16.3 DO Calibration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146
16.3.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151
16.4 Calibrating the Turbidity Sensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152
16.4.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153
16.5 Redox Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154
16.5.1 Functional Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
16.5.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
16.6 Totalizer for Pumps and Valves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
16.6.1 Pump Calibration Sequence . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
16.6.2 Scale/Balance Calibration Sequence . . . . . . . . . . . . . . . . . . . . . 161
17. “Controller” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
17.1 Functional Principle and Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
17.2 Controller Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
17.3 General Controller Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
17.4 Setpoint profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
17.4.1 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168
17.4.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168
17.5 General Controller Parameterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168
17.5.1 Output Limits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
17.5.2 Dead Zone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
17.5.3 Controller Parameterization Menu Screen . . . . . . . . . . . . . . . . 170
17.5.4 PID Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170
17.5.5 PID Controller Optimization . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170
17.6 Temperature controller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
17.6.1 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172
17.7 Speed Regulator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172
17.7.1 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 173
17.8 pH Controller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174
17.8.1 Operating Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
17.8.2 DO Supply-driven pH Control . . . . . . . . . . . . . . . . . . . . . . . . . . 175
17.8.3 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
17.9 DO Control Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
17.9.1 pO2 Controller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
17.9.2 Advanced DO Controller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179
17.9.3 Parameterizing the Master Controller . . . . . . . . . . . . . . . . . . . . 179
17.9.4 Selection and Configuration of the Slave Controllers . . . . . . . 184
17.9.5 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
17.9.6 Instructions for Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
17.10 Gas Filler Controller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
17.10.1 Operating Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
17.10.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
6 Contents
17.10.3 Gas Flow Controller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
17.11 Foam and Level Controllers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194
17.11.1 Displays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
17.11.2 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
17.11.3 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
17.12 Gravimetric Filling Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196
17.12.1 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196
17.12.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196
17.13 Filling Pump Controller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197
17.13.1 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197
17.14 Pump Assignment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 198
17.14.1 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199
17.14.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199
18. “Phases” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 200
18.2 Phase Process Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201
18.2.1 Status Displays During Step Control . . . . . . . . . . . . . . . . . . . . . 202
18.2.2 General Procedure for Phase Control . . . . . . . . . . . . . . . . . . . . . 203
18.2.3 Displaying Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204
18.2.4 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 206
18.3 Sterilization Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 206
18.4 Additional Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
18.4.1 Pressure Hold Test of Culture Vessel . . . . . . . . . . . . . . . . . . . . . . 208
19. “Settings” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 209
19.2 System Settings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210
19.3 Measuring Range Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 211
19.4 Manual Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
19.4.1 Manual Operation of Digital Inputs . . . . . . . . . . . . . . . . . . . . . . 214
19.4.2 Manual Operation of Digital Outputs . . . . . . . . . . . . . . . . . . . . 215
19.4.3 Manual Operation of Analog Inputs. . . . . . . . . . . . . . . . . . . . . . 218
19.4.4 Manual Operation of Analog Outputs . . . . . . . . . . . . . . . . . . . . 220
19.4.5 Manual Operation for Controllers (”Control Loops”) . . . . . . . . 221
19.4.6 Manual Operation for Counters (”Digital Counters”) . . . . . . . . 223
19.4.7 Manual Operation of Sequence Control (”Phases”) . . . . . . . . . 224
19.5 Externally Connected Devices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225
19.6 Service and Diagnosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
20. Appendix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
20.1 Alarms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
20.1.1 Alarm Triggering. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
20.1.2 “Alarm Overview” Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228
20.2 Process Value Alarms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228
20.2.1 Operating Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 230
20.2.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 230
20.3 Alarms for Digital Inputs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231
20.3.1 Operating Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232
20.3.2 Special Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232
20.4 Alarms, Meaning and Corrective Measures . . . . . . . . . . . . . . . . . . . . . . . 232
20.4.1 Process Alarms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232
20.4.2 Process Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
20.4.3 System Alarms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
20.5 Bug Handling and Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
20.6 Locking Functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
20.7 GNU Licensing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
20.8 Password System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 234
Contents 7
8
Part A: BIOSTAT® Cplus
Operating Manual
(Original Operating Manual)
Fermenter | Bioreactor
9
1. Introduction
1. Introduction
All information and instructions in this operating manual have been compiled in
consideration of applicable standards and guidelines, the state of the art, and our
many years of experience and knowledge.
In addition to the operating manual, all generally valid, legal and otherwise binding
regulations for accident protection and environmental protection of the country of
use must be observed.
This operating manual provides you with all the information you need for installing
and operating the BIOSTAT ® Cplus unit (called the “device” in this document).
t Read the operating manual carefully before carrying out processes on the device.
t This manual is part of the product. Keep it in a safe and easily accessible place near
the device’s site of installation.
t If the manual should be lost or misplaced, please contact Sartorius for a

replacement or download the latest version from our website: www.sartorius.com
The operating manual applies to the BIOSTAT® Cplus in combination with the
following culture vessels:
− Stainless steel culture vessel, double-walled, with upper glass shot
(working volume):
− 5L
− Stainless steel culture vessel, double-walled (working volume):
− 10 L
− 15 L
− 20 L
− 30 L
The device may only be used with the equipment and under the operating
conditions described in the Technical Data Sheet [¨ Chapter “10.1
Specifications”].
The user must be qualified to handle the device, media and cultures and be
aware of the hazards potentially associated with the process.
The process may make it necessary to equip the device or the work area with
additional safety equipment or to take other measures for the protection of
personnel and the work environment.
The documentation does not include more detail about such conditions or legal
or otherwise obligatory guidelines.
Safety and danger instructions in the documentation only apply to this device
and supplement the guidelines of the operator in the work area for the process
in question.
The model designation can be found on the model plate. The model plate is located
on the control/supply device and on the culture vessel.
See also [¨ Section “5.1.5.2 Type Plates”].
The operating instructions must be read, understood, and used by all personnel
entrusted with the operation, service, cleaning, and troubleshooting of the device.
This applies particularly to the safety instructions listed.
10 Introduction
After reading the operating manual, you will be able to
− Operate the device safely,
− Service the device according to guidelines,
− Clean the device according to guidelines,
− Take appropriate measures should a fault occur.
1.1 Presentation As a means of instruction and direct warning of danger, all text statements to be
particularly noted in this operating manual will be marked as follows:
This symbol denotes an immediate danger with high risk that death or (severe)
injury may result if it is not avoided.
This safety instruction denotes a possible danger with medium risk that death or
(severe) injury may result if it is not avoided.
This symbol denotes a possible danger with risk that moderate or minor injury
may result if it is not avoided.
This symbol denotes a danger with the low risk that could result in property
damage if the risk is not avoided.
This symbol
− is an indication of a function or setting on the device,
− or that caution should be exercised while working
− or identifies useful information.
The following presentations will also be used:
− Texts that follow this mark are lists.
t Texts that follow this mark describe activities that must be carried out in the
specified order.
y Texts that follow this mark describe the result of an action.
1. Texts that follow this mark describe activities that must be carried out in the
numbered order.
” “ Texts in quotes are references to other chapters or sections.
[¨ ] Texts following this symbol make reference to other chapters, sections or

documents.
Introduction 11
1.2 Service Center Repairs can be carried out on-site by authorized service personnel or by the
responsible Service representative of Sartorius Stedim Systems GmbH.
The model designation can be found on the model plate or signage [¨ see Section
“5.1.5.2 Type Plates”].
For equipping and retrofitting as well as repairs, you may only use parts approved by
Sartorius Stedim Systems GmbH for the unit.
Sartorius Stedim Systems GmbH shall not be liable for repairs made by the customer
or any resulting damage.
The warranty shall expire particularly in the case of:
− The use of unsuitable parts that deviate from the device’s specifications.
− Modification to parts without the approval of Sartorius Stedim Systems GmbH.
In the event of service requests or warranty claims, please notify your representative
of Sartorius Stedim Systems GmbH and/or Sartorius Stedim Biotech GmbH or contact:
Sartorius Stedim Systems GmbH

Robert-Bosch-Str. 5–7
D-34302 Guxhagen, Germany
Tel. no. +49 (0) 5665 407-0
Fax. no. +49 (0) 5665 407-2200
E-Mail: info@sartorius-stedim.com
Website: http://www.sartorius-stedim.com
Returning Devices
Send defective devices or parts to Sartorius Stedim Systems GmbH.
Returned devices must be clean and in hygienically flawless condition and packed
carefully. Contaminated parts must be disinfected and/or sterilized in accordance
with the safety guidelines applicable to the area of application.
The sender must prove compliance with the regulations. To do so, use the
decontamination declaration in the appendix [¨ Section “10.1 Specifications”].
Transport damage, as well as measures for subsequent cleaning and disinfection of

the parts by Sartorius Stedim Systems GmbH, shall be charged to sender.
12 Introduction
2. Safety Instructions
2. Safety Instructions
This document only contains safety information on dangers when handling the
devices and possible preventive measures. It does not go into detail on process-
dependent risks and applicable legal or otherwise obligatory guidelines for the
protection of personnel and working environment.
Failure to observe the following safety precautions can have severe

consequences:
− Danger to personnel due to electrical, mechanical, or chemical influences
− Failure of important device functions
Read the safety precautions and dangers listed in this section thoroughly before
starting the device.
The operator | user must observe all existing national workplace, operating, and
safety guidelines in addition to the instructions in this operating manual.
Existing internal plant guidelines must also be observed.
2.1 General Safety − The device may only be started and serviced after this operating manual has been
Instructions read and understood by the operators.
− Use the device only for its intended purpose [¨ Chapter “2.4 Intended Use and
Foreseeable Misuse”].
− The device is not ATEX (ATmosphère EXplosive)-certified. The device may not be
operated in potentially explosive atmospheres.
− During operation of the device, do not permit any work method that hinders the
safety of the device.
− Always keep the working area of the device clean and orderly, in order to avoid
danger from dirt or scattered parts.
− Always squat to work on low-lying components; do not bend over. Carry out work
on high components in an upright, straight body position.
− Do not exceed the technical performance data for the device
[¨ see Section “10.1 Specifications”].
− The device may only be used indoors.
− Keep all safety precautions and danger descriptions at the device in legible
condition, and replace them as needed.
− Operation as well as work on the device may only be carried out by trained
personnel.
− Do not start the device if other people are in the danger zone.
− Chemicals that can attack housing, housing gaskets and cable sheathings must be
kept away from the device.
These include oil, vegetable and animal fats, petrol, chlorinated and aromatic
solvents, lye and acids, acetone and ozone. If you are uncertain, contact the
manufacturer.
The casing on all connection cables as well as the wires inside the equipment
housing are made of PVC. Chemicals that corrode this material must be kept away
from these cables.
− In case of malfunction, immediately stop the device.
Have the fault corrected by appropriately trained personnel or by your Sartorius
Stedim Service office.
Safety Instructions 13
2.2 Informal Safety Measures − Always keep the operating manual in the location where the device is in use.
− In addition to the operating manual, follow all general and local regulations for
accident prevention and environmental protection.
2.3 Symbols Used The following symbols are used on the device:
on the Device
Especially hazardous area or hazardous use!
Follow the instructions in the accompanying documents.
Danger of crushing your hand or fingers in the peristaltic pump – do not

attempt to put your hand inside rotating parts, e.g. the pump head!
To thread the tubing or adjust the rollers, be sure to
switch off the pump first.
Danger of burns!
Motor and culture vessel equipment becomes hot during operation.
− Avoid accidental, unintentional contact.
− Use protective gloves when operating the equipment.
− Prior to removing the motor from the stirrer drive, allow the motor housing to cool
down.
− Let the culture vessel and equipment cool down before carrying out assembly
work.
Equipment manufacturers have placed some identifying marks, symbols or labels on

the various pieces of equipment. These may not always correspond to the safety labels
commonly used by Sartorius Stedim Systems.
Be sure to follow the instructions given in this manual.
− Keep all safety precautions and danger descriptions at the device in legible
condition, and replace them as needed.
2.4 Intended Use and The operational safety of the device is only ensured when it is used for its intended
Foreseeable Misuse use and operated by trained personnel.
The device is used for the cultivation of prokaryotic and eukaryotic cells in aqueous
solution.
Only biological materials in Groups 1 and 2 may be used in the device.
The intended use also includes:

− The observation of all instructions from the operating manual.
− Observation of inspection and maintenance intervals.
− The use of oils and greases which are suitable for use with oxygen.
− The use of operating and auxiliary materials according to applicable safety
guidelines.
− Observation of operating and repair conditions.
All other applications are not considered the intended use. They may include risks
that cannot be estimated and they are the sole responsibility of the operator.
14 Safety Instructions
Claims of any kind resulting from damages due to use other than the intended use are
excluded.
Sartorius Stedim Systems GmbH bears no liability for use other than the intended use.
Danger due to use other than the intended use!

Any use beyond and/or other than the intended use of the device can lead to
dangerous situations.
The following uses are considered not the intended use and are strictly forbidden:
− Processes using biological materials in Safety Classes 3 and 4
− Cultivation in non-aqueous solutions
− Overloading the device
− Work with parts under voltage
− Operation outdoors
2.5 Residual Risks from This device is a state-of-the-art machine and is built in accordance with recognized
Use of the Device safety standards. Nevertheless, the use of the device may pose bodily or physical harm
to users or third-parties as well as potentially cause impairments to the testing
system itself or other material damage.
Anyone assigned to the installation, commissioning, operation, maintenance or repair

of the device must have read and understood the operating manual.
The device may only be used:

− for its intended use.
− with its safety systems in perfect working order.
− by qualified and authorized personnel.
Furthermore, the following rules must be followed:

− All moving parts must be lubricated as required.
− All screw connections must be checked at regular intervals and tightened if
necessary.
During use of the device, the following residual risks still occur:
− During assembly and disassembly of vessel components (cover, sampling container,
feed mechanisms) there is a risk of impact or crushing.
− Rotating parts (agitator)
− Danger of burns due to hot surfaces
2.6 Danger Due to
Danger to life caused by electrical voltage!
Electrical Power
Electrical switching elements are installed in the device. Contact with parts under
voltage represents a direct danger of death. Damage to the insulation or individual
components can be fatally dangerous.
− Never open the device. The device may only be opened by authorized personnel of
the Sartorius Stedim Biotech Company.
− Work on the electrical equipment of the device may only be carried out by
Sartorius Stedim Service or authorized technicians.
− Check the electrical equipment of the device regularly for defects such as loose
connections or damage to the insulation.
− In case of defects, turn the power supply off immediately and have the defects
corrected by Sartorius Stedim Service or authorized technicians.
− If work is required on parts under voltage, ask a second person to be ready to turn
off the device's main switch if needed.
− During all work on the electrical equipment, disconnect it and check that voltage
is no longer applied.
− During maintenance, cleaning, and repair work, turn the power supply off and
secure it against reactivation.
− Keep moisture away from parts under voltage, as it can lead to short circuits.
− Electric power lines may not be placed on hot pipes.
− Have the electrical components and stationary electrical equipment checked by an
electrician at least every 4 years.
− Have non-stationary electric equipment, cables with plugs, and extension or
connector cables and their connectors, if any, checked at least every 6 months by
an electrician or, if using suitable testers, by a trained person.
Non-stationary equipment is any equipment that is designed to be continually moved

while still connected to power. This includes electrical floor cleaning systems, for
example.
2.7 Danger Due to

Danger of injury from leaked substances!
Components Under
If individual components are damaged, gaseous and liquid materials may escape
Pressure
under high pressure and cause injury to eyes, for example.
Therefore:
− Do not start the culture vessel without a safety valve or comparable overpressure
safety (e.g. a burst disk).
− Turn the device off and secure it against reactivation when working on it.
− Release the pressure from system sections and pressure lines to be opened before
starting any repair work.
− Regularly check all lines, hoses, and connections under pressure for leaks and
externally detectable damage.
2.8 Hazards Arising from
Danger of injury from shattered glass!
Bursting Culture Vessel
Damaged and bursting culture vessels can cause cuts and
eye injuries.
Therefore:
− Provide training for your personnel with regard to glass breakage due to outside
effects.
− Do not sterilize the 5 L culture vessel without a protective jacket.
− Wear personal protective equipment.
− Ensure that the culture vessel is securely connected to the supply and control unit.
− Ensure that the culture vessel remains within the maximum permissible pressure
limits.
− Regularly check all lines, hoses, and connections under pressure for leaks and
externally detectable damage.
2.9 Danger Due to Gases
2.9.1 Dangers Due to

Danger of explosion and fire!
Oxygen
− Keep pure oxygen away from flammable materials.
− Avoid sparks in the vicinity of pure oxygen.
− Keep pure oxygen away from ignition sources.
− Keep the entire aeration segment oil- and grease-free. Ensure that cooling water
flowing back is unpressurized.
− Monitor the limit values at the system and in the building.
Reaction with other materials!

− Ensure that oxygen does not come into contact with oil or grease.
− Use only materials and substances suitable for use with pure oxygen.
2.9.2 Danger Due to Danger of suffocation due to escaping nitrogen!

Nitrogen
Escaping gas at high concentrations may force the air out of closed rooms
and can cause loss of consciousness, leading to suffocation.
− Check the gas lines and culture vessels for leaks.
− Provide good ventilation where the device is installed.
− Keep a breathing device independent of ambient air ready for emergencies.
− If personnel appear affected by symptoms of suffocation, immediately provide
them with a breathing device independent of ambient air,
− bring them into fresh air, make them comfortable, and keep them warm. Call a
doctor.
− If a person stops breathing, initiate first aid measures with artificial respiration.
− Check process gas lines and filters regularly for leaks.
2.9.3 Danger Due to

Danger of poisoning by escaping carbon dioxide!
Carbon Dioxide
− Check the gas lines and culture vessels for leaks.
2.10 Danger Due to
Danger of scalding due to defective components!
Escaping Steam
− Inspect the device before starting the process.
− Check the connections of containers and the connections to the supply unit.
− Regularly check hoses for leaky places and replace any leaking hoses.
2.11 Danger Due to Danger of injury from escaping feed and culture media!
Escaping Materials − Use the specified hoses only.
− Use hose fastenings on connecting pieces.
− Empty the feed hoses before loosening the hose connection.
− Take care that hoses are laid without kinking or pinching.
− Wear safety glasses.
Danger of contamination from escaping feed and culture media!

− Empty the feed hoses before loosening the hose connection.
− Wear safety glasses.
2.12 Danger Due to

Danger of burns upon contact with hot surfaces!
Hot Surfaces
− Avoid contact with hot surfaces like culture vessels, motor housings, and pipelines
carrying steam.
− Block the danger zone off.
− Wear protective gloves when working with hot culture media.
− Electric power lines may not be placed on hot pipes.
2.13 Danger Due to
Danger of limbs being pulled into the rotation pump and crushed!
Rotating Parts
− Do not remove the safety mechanisms.
− Allow only qualified and authorized personnel to work on the device.
− Disconnect the device from power when performing maintenance and
− cleaning tasks.
2.14 Danger Due to

Danger of injury due to incorrect consumables!
Use of Incorrect
− Incorrect or erroneous consumables can lead to damage, malfunction, or the total
Consumables
failure of the device, as well as affecting safety.
− Use only original consumables.
Purchase consumables through Sartorius Stedim Service GmbH. You can find all
necessary specifications for consumable materials in the overall documentation.
2.15 Personal Protective When operating the device, personal protective equipment must be worn in order to
Equipment minimize dangers to health.
− During work, always wear the protective equipment needed for that work.
− Follow any instructions posted in the work area pertaining to personal protective
equipment.
Wear the following personal protective

equipment during all work:
Protective work clothes

Protective work clothing is tight-fitting work clothing with low tear resistance, tight
sleeves, and without any projecting parts. It is primarily intended to protect you from
being caught by moving machine parts.
Do not wear rings, chains, or other jewelry.
Head covering
To protect your hair from being caught and pulled into moving parts of the device,
wear a head covering.
Safety gloves
Wear safety gloves to protect your hands from process materials.
Safety glasses
Wear safety glasses to protect yourself from media escaping under high pressure.
Safety boots
Wear non-slipping safety boots to protect against slipping on smooth floors.
2.16 Safety and Protective
Systems
2.16.1 Switch-Disconnector Control Unit
The switch-disconnector is on the left side of the control unit's housing and is
designed to be a physical on | off switch for the power supply. The switch-
1
disconnector is simultaneously the main switch used to turn the device on and off.
2.16.2 Safety Valves and Danger of injury from bursting culture vessels and lines!
Pressure Reducer − Do not start the device without safety valves and pressure reducer or comparable
overpressure safety (e.g. a burst disk).
− Have your safety valves and the pressure reducer serviced regularly by the
Sartorius Stedim Service and replace broken burst disks immediately.
− Follow the information in the overall documentation.
Safety Valve / Burst Disk

The safety valve or burst disk is a component of the vessel's equipment [¨ see
Chapter “3. Device Overview”].
− The safety valve is installed on the lid (PED and SELO culture vessels).
− The burst disk is installed in the upper region of the vessel wall (ASME culture
vessels).
In addition, there is a safety valve in the temperature control circulation.
The safety valve or burst disk triggers at a defined pressure.

This prevents impermissible overpressure, ensuring safe operation.
2.17 Emergency Instructions Preventive Measures

− Always be prepared for accidents or fire.
− Keep first aid equipment (bandages, blankets, etc.) and fire extinguishers handy.
− Familiarize personnel with accident reporting, first aid, fire extinguishing, and
rescue facilities.
− Keep entry and rescue routes free for rescue vehicles and rescue personnel.
Measures To be Taken After Accidents
− Trigger an emergency stop at the switch disconnector.
− Keep personnel out of the danger zone.
− In case of stopped heart | or breathing, initiate first aid measures immediately.
− Report personal injuries to the first aid officer and an emergency doctor and | or
the rescue service.
− Keep entry and rescue routes free for rescue vehicles and rescue personnel.
− Extinguish fires in the electrical equipment with a CO2 extinguisher.
2.18 Operator Responsibilities The device is used in the commercial sphere. The operator of the device is therefore
subject to the legal obligations for workplace safety.
In addition to the safety instructions in this operating manual, the safety, accident
prevention, and environmental protection regulations valid for the location of use of
the device must also be observed.
In particular, the following apply:

− The operators must inform themselves of the applicable workplace safety
regulations and carry out a hazard evaluation to determine additional dangers
that may result due to the special working conditions at the location of use of the
device. These must be expressed in the form of operational instructions for the
operation of the device (hazard prevention plan).
− During the entire period of use of the device, the operator must check whether the
operating instructions correspond to the current state of regulations and adapt
them as needed.
− The operator must clearly regulate and determine responsibilities for operation,
maintenance, and cleaning.
− The operator may only allow trained and authorized individuals to work with the
device. Trainees, apprentices and auxiliary staff members may only work with the
device under the supervision of specialist personnel [¨ Section “2.19 Personnel
Requirements”].
− The operator must ensure that all employees involved with the device are suitable
in terms of physical capacity, person and character to operate the device
responsibly.
− The operator must also ensure that all employees are familiar with the basic
regulations concerning workplace safety and accident prevention, trained in
handling the device, and have read and understood the operating manual.
− Furthermore, the operator must evaluate the safety-conscious work of personnel
at regular intervals and demonstrate training of personnel and notification of
danger.
− The operator must avoid stress situations during operation of the device by means
of technological and organizational work preparation.
− The operator must provide adequate workplace lighting in the place of operation
of the device in accordance with locally applicable workplace guidelines.
− The operator must provide personnel with personal protective equipment.
− The operator must ensure that no-one works on the device whose reaction time is
impaired, for example by drugs, alcohol, medications, or the like.
Furthermore, the operator is responsible for ensuring that the device is always in
technically perfect condition.
The following therefore applies:
− The operator must ensure that the maintenance intervals described in this
operating manual are observed.
− The operator must regularly have the safety systems tested for functionality.
2.19 Personnel
Danger of injury if qualifications are insufficient!
Requirements
Improper use can lead to significant personal injury and | or property damage.
Have all activities performed only by qualified personnel.
Only those people are permitted as personnel of whom it can be expected that they
will carry out their work reliably. No-one may work on the device whose reaction
time is impaired, for example by drugs, alcohol, medications, or the like.
2.20 Personnel Qualification In the operating manual, the following qualifications are cited for different areas of
Requirements activity:
Trainee
A trainee such as an apprentice or an auxiliary staff member does not know all the
dangers that can occur during operation of the device. They may only perform work
on the device under the supervision of technicians.
Trained Person
A trained person has been informed in a training session by the operator about the
tasks assigned to them and the possible dangers of improper behavior.
Technician
Due to his or her technical education, knowledge, and experience, as well as
knowledge of applicable regulations, a technician is capable of carrying out tasks
assigned to him or her and independently detecting and avoiding possible dangers.
Electrician
Due to his or her technical education, knowledge, and experience, as well as
knowledge of applicable standards and regulations, an electrician is capable of
carrying out work on electrical equipment and independently detecting and avoiding
possible dangers.
The electrician is trained for the special place of use where he or she works and knows
the relevant standards and regulations.
2.21 Personnel Responsibilities Before undertaking any work with the device, all personnel are obliged to:
− Pay attention to the basic occupational safety and accident prevention
regulations.
− Read the safety instructions and warnings in this operating manual and confirm
by signature that they are understood.
− Follow all safety and operation instructions in this operating manual.
2.22 Responsibilities The responsibilities of personnel for the operation, maintenance, and cleaning must
be clearly defined.
2.23 Unauthorized
Danger to unauthorized personnel!
Personnel
Unauthorized personnel who do not meet the qualification requirements for
personnel do not know the dangers in the work area.
Therefore:
− Keep unauthorized personnel away from the work area.
− In case of doubt, speak to personnel and instruct them to leave the work area.
− Stop work as long as unauthorized personnel remain in the work area.
2.24 Training and Instruction Personnel must regularly receive instruction from the operator.
Log the performance of this training for better tracking.
Date Name Type of Training Training Given by Signature
3. Device Overview
3. Device Overview
The BIOSTAT® Cplus bioreactor is designed for cultivating microorganisms and cells in
discontinuous and continuous processes.
Vessels are available with working volumes of 5, 10, 15, 20, and 30 liters. They can
have a height | diameter ratio of 2:1 or 3:1.
The device is made up of the following components:

− Control unit with DCU control system and integrated pumps and aeration system.
− Supply unit as a frame construction with temperature control system and culture
vessel mount.
− Culture vessel with equipment and attachments.
The BIOSTAT® Cplus with 5 L culture vessel is a bench-top system.

The BIOSTAT® Cplus with 10 to 30 L culture vessel is a floor-standing system.
This chapter describes the standard vessel and equipment for the BIOSTAT® Cplus
bioreactor.
The illustrations in the following sections show some example system configurations.
The actual equipment depends on your configuration and may deviate from the
bioreactors shown here.
26 Device Overview
3.1 Overall Views The following illustrations show an example of the BIOSTAT® Cplus bioreactor with its
control unit.
Overview BIOSTAT® Cplus using the example of the Cplus L (floor-standing)
11
8
4
5
9
6
1
10
Pos. Description
1 Control unit with pumps and aeration module
2 Frame
3 Culture vessel
4 Drive motor
5 Supply air filter
6 Exhaust cooler
7 High foam adapter (optional)
8 Exhaust air filter
9 Culture vessel safety valve
10 Floor drain valve
11 Pressure control valve (optional)
Device Overview 27
Overview BIOSTAT® Cplus using the example of the Cplus C-15-3
(floor-standing)
Pos. Description
1 Control unit with pumps and aeration module
2 Frame
3 Culture vessel
4 Drive motor
5 Supply air filter
6 Exhaust cooler
7 High foam adapter (optional)
8 Exhaust air filter
9 Culture vessel safety valve (not available on ASME culture vessels)
9a Burst disk with vent duct (only ASME culture vessels, not pictured)
10 Floor drain valve
11 Motor mount (optional) with motor
12 Corrective solution bottles with holder
13 Steam distributor (not pictured)
28 Device Overview
3.2 Control | Supply Units Front view
3
4
5
Fig. 3-1: BIOSTAT® Cplus control unit - Advanced Additive Flow version
Pos. Description
1 Operator terminal (touch panel)
2 Operating state lamp
Lamp is lit: Device in service
Lamp not lit: Device out of service
3 Headspace aeration “Overlay”
4 Medium aeration “Sparger”
5 Variable area flow meter (rotameter)
6 Peristaltic pumps
Device Overview 29
Rear view
1 2 3 4
Fig. 3-2: Control unit connection panel, reverse side (in the lab)
Pos. Description
1 “AIR” laboratory connection, quick-connect coupling d 6 mm
2 “O2” laboratory connection, quick-connect coupling d 6 mm
3 “N2” laboratory connection, quick-connect coupling d 6 mm
4 “CO2” laboratory connection, quick-connect coupling d 6 mm
30 Device Overview
Side view
3
4
5
Fig. 3-3: Control unit connection panel, side
Pos. Description
1 DO (“pO2”): DO sensor, VP8 plug
pH: pH sensor VP8 plug
Temp: Temperature sensor Pt-100, M12 female connector
Pressure: Pressure sensor, M12 female connector
2 Turbidity: Sensor for turbidity measurement, Lemo socket
Lamp: Lamp connector for viewing window, Amphenol socket
Level: Level sensor, M12 female connector
High Foam: “High Foam” sensor, M12 female connector
Foam: “Foam” antifoam sensor, M12 female connector
3 Serial-A: Scale connection, M12 female connector
Serial-B: Scale connection, M12 female connector
Pump-C: External pump connection, M12 female connector
Pump-D: External pump connection, M12 female connector
4 Main switch / switch-disconnector “Main”
5 Ethernet: Network interface, M12 female connector
Ext. sig. A | B: External signal output, M12 female connector
Ext. sig. C: External signal output, M12 female connector
Device Overview 31
6
10
11
Fig. 3-4: Control unit connection panel, side (continued)
Pos. Description
6 Headspace aeration “Overlay”, tube connector d 6 mm
7 Medium aeration “Sparger”, tube connector d 6 mm
8 Autom. valves: Supply unit valve connections, Amphenol female connector
Pressure: Pressure control valve connection, M12 female connector
Balance: Tank weighing connection, M12 female connector
J. Temp: Temperature sensor in the temperature control circuit, M12 female
connector
9 Electrical Heater: Electric heating connection, Amphenol female connector
Circulation Pump: Pump in the temperature control circuit, Amphenol female
connector
10 Power supply connection:
400 V, CEE plug, 5-pin
208 V NEMA L21-20P plug, 5-pin
11 Manufacturer's ID label, supply | control unit
32 Device Overview
3.3 Aeration The control unit of the BIOSTAT® Cplus can be equipped with a range of different
aeration modules. Every control unit can only be equipped with one of the aeration
module models described below.
− Please observe the P&I diagram in addition to the aeration module specifications
of the bioreactor.
Variable area flow meter

All of the aeration modules are fitted with a variable area flow meter. The ranges of
the measurement and control equipment supplied with the aeration modules will
match the culture vessel they are intended for or will be as agreed in the order
confirmation. On request, aeration module measurement and control equipment can
also be changed after delivery.
Mass flow controllers must be ordered separately, i.e. They will be included in the
standard equipment supplied for the bioreactor only if you specified these on your
order.
If fitted retrospectively, they must only be fitted by an authorized service technician.
− Mass flow controllers must be regularly recalibrated as specified by the
manufacturer [ ¨ Manufacturer's documentation].
− Recalibration work must only be performed by authorized service technicians or
the manufacturer. Please refer to the appendix for more information on sending
items to Sartorius Stedim Systems GmbH.
− If the gas flow of a gas or the total aeration rate is controlled via built-in mass
flow controllers, completely open the variable area flow meter when no gas flow
limitation shall pass through.
Designations on the variable area flow meter

To identify the variable area flow meters, their aeration module is labeled.
The following nomenclature is used:
Description Meaning
Air Air
O2 Oxygen
N2 Nitrogen
CO2 Carbon dioxide
Supplemetal Information
The built-in variable area flow meters are calibrated to the following standard
conditions.
Calibration parameters
Gas type: Air
Temperature: 20° C = 293 K
Pressure: 4 bar (absolute)
When gases with deviating pressures pass through, higher or lower values can be
displayed. These must be recalculated to evaluate the flow rates.
The manufacturer of the flow rate meters provides tables with conversion factors.
Using the conversion tables, flow rates for the different processes can be recalculated.
Device Overview 33
Specific Data for Gas Density [kg/m3]
Carbon dioxide (CO2) 1.977
Air 1.293
Oxygen (O2) 1.429
Nitrogen (N2) 1.251
3.3.1 “O2 Enrichment” and The “O2 Enrichment” and “Gasflow Ratio” aeration modules supply air and enriches it
“Gasflow Ratio” with oxygen, e.g. for microbial cultures.
Aeration
Fig. 3-5: Aeration module “O2 Enrichment” and “Gasflow Ratio”, with outlet “Sparger”
“O2 Enrichment” Aeration

In the “O2 Enrichment” aeration module, aeration takes place with “Air” as the carrier
stream, which may be enriched with oxygen via a solenoid valve.
The total aeration rate is configured on the variable area flow meter; optionally, this
can be controlled by a mass flow controller.
− Oxygen enrichment takes place via a 3/2-way solenoid valve, the solenoid valve is
controlled via the DCU system's gas filling controller/DO controller.
− The mass flow controller is controlled via the DCU system's gas filling controller/
DO controller.
− Output “sparger” for culture vessel total aeration rate.
“Gasflow Ratio” Aeration

In the “Gasflow Ratio” aeration module, aeration takes place with “Air” and/or with
oxygen, each with a mass flow controller.
− The mass flow controller is controlled via the DCU system's gas filling controller/
DO controller.
34 Device Overview
Aeration “Advanced Additive Flow - 2 out”
Fig. 3-6: Aeration “Advanced Additive Flow - 2 out”
In the “Advanced Additive Flow - 2 out” aeration module, aeration takes place with
up to 4 gases via a “sparger” and an “overlay” output. These are by default:
− Supply of air for “Sparger” and “Overlay”
− N2 for decreasing the O2 content, or
O2 for the “Sparger” for increasing the oxygen content;
− CO2 for adjusting the pH or use as a carbon (C) source for “Sparger”
The aeration rates are set at variable area flow meters.

Optionally, these can be controlled by mass flow controllers.
− Oxygen enrichment / depletion takes place via 2/2-way solenoid valves or mass
flow controllers.
The solenoid valve / mass flow controller is controlled by gas filling controllers /
gas flow controller or the DCU system's DO controller.
− CO2 can be added either via a 2/2-way solenoid valve or a mass flow controller.
The solenoid valve / mass flow controller is controlled by gas filling controllers /
gas flow controllers or the DCU system's pH controller.
Device Overview 35
3.4 Pump Modules
The integrated hose pumps are designed for use with silicone hoses. Other tubing
materials can significantly shorten the life of the hose pump.
The peristaltic pumps are located on the supply unit and convey the correction media
and nutrient media through hoses into the vessel.
Up to 4 peristaltic pumps can be installed per control unit.
Using soft keys on the control unit, the pump can be switched on to fill the tubes, for
example. In addition, the pumps are automatically controlled in the respective control
loops in the operating mode “Auto”.
The pumps are labeled with stickers according to their standard function.
Fig. 3-7: Integrated peristaltic pumps
3.4.1 Performance The peristaltic pumps can be installed in the supply unit in 2 different specifications:
Parameters and
Features
Type Speed (rpms)
WM 102 fixed speed 20 rpm

WM 102 regulated speed 5 - 50 rpm
The following overview shows the possible silicone hoses with the feed rate per
revolution:
Tubing wall thickness 1.6 mm

Inner diameter of the hose (mm) 0.5 0.8 1.6* 3.2 4.8
Flow rate: ml/revolution 0.02 0.05 0.22 0.81 1.66
* = tubing sizes delivered by default
36 Device Overview
3.4.2 External Pumps External pumps can be connected to the supply unit. See [¨ Section “3.2 Control |
Supply Units“] for the connections.
3.5 Culture Vessels Culture vessels are available with operating volumes of 5, 10, 15, 20 and 30 liters.
Culture vessels 10, 15, 20 and 30 liters

The 10-, 15-, 20- and 30-liter culture vessels may have a height / diameter ratio of
2:1 or 3:1 and consist of a solid steel vessel.
Culture vessel 5 Liter

The 5-liter culture vessel has a height / diameter ratio of 2:1 and consists of a glass
cylinder with a stainless steel base part.
All culture vessels have a double outer wall for temperature control. The double wall
and other vessel attachments, such as the floor drain valve and the SVC 25 sampling
valve are connected to the frame installation via flexible connection lines braided
with stainless steel. Additional connections are designed as flexible tubing
connections.
Device Overview 37
3.5.1 Overview, 5-liter
Culture Vessel
Culture Vessel CT 5-2, from the Outside Culture Vessel CT 5-2, Internal Construction
Pos. Description Pos. Description Pos. Description

1 Vessel Bottom, Double Wall 9 Floor Drain Valve 36 Glass Cylinder
2 Cover Plate 18 Rotating Mechanical Seal 37 Screw Connections
3 Sparger Pipe (Ring Sparger) 22 Stirrer Shaft Coupling 41 Rotating Mechanical Seal
4 Gas Inlet, with N12 Filter 23 Gasket, Glass Cylinder 44 Screws on the Ring Flange (6x)
5 Exhaust Air Filter 31 Bearing Housing, Stirrer Shaft 43 Splinter Protection Sheath
6 Impeller 33 Lid Screw Connections (4x) 66 Assembly “High Foam Alarm”
8 Exhaust Cooler 35 Ring Flange, Glass Cylinder
38 Device Overview
3.5.1.1 Connections in the
Cover Plate
Fig. 3-8: Cover plate design Cplus 5-2
Pos. Description Placement, mounting, other comments

1 Level probe Cover port d = 19 mm
2 Safety valve Cover port d = 15 mm
3 Stirrer shaft connectionCover port d = 45 mm; single mechanical seal or
double mechanical seal
4 Exhaust cooler Cover port d = 30 mm
5 Reserve port Cover port d = 19 mm; e.g. for inoculation fittings
6 Locking bolts 4 locking bolts for securing the cover plate
7 Reserve port Cover port d = 19 mm; e.g. for harvest fittings
8 Supply air filter Cover port d = 19 mm,
with adapter for in-situ sterilization
9 Condensate intake/drain Only for double mechanical seal
10 Antifoam probe Cover port d = 19 mm
3.5.1.2 Side Ports on the

Culture Vessel
Culture vessel wall

Pos. Description
1 Port d = 12 mm, level, for Pt 100
2 Port d = 25 mm, slanted, for installation of the pH or DO electrode or as a
reserve
3 Port d = 25 mm, slanted, as in (2)
4 Port d = 25 mm, level, for SVC 25
5 Port d = 25 mm, slanted, as in (2)
Culture vessel bottom

1 Port d 32 mm for floor drain valve
2 Port R 3/8“ connection of the double wall to the thermostat system
Device Overview 39
3.5.2 Overview, 10-liter to
30-liter Culture Vessel
3.5.2.1 Connections in the

Cover Plate 13
1
12
11 2
3
10 4
9 5
8
6
7
Fig. 3-9: Cover plate design Cplus
Pos. Description Placement, Mounting, other Comments

1 Antifoam Probe Cover Port d = 19 mm
2 Exhaust Cooler Cover Port d = 30 mm
3 Stirrer Shaft Coupling With single or optional double mechanical seal;
installed in cover port d = 45 mm
4 Cover Viewing Window Cover port d = 25 mm equipped with viewing window for illumination.
5 Reserve Port Cover port d = 19 mm; e.g. for harvesting pipe
6 Lid Screw Connection Securing the cover plate with 4-6 locking bolts or hexagon nuts (depending on culture vessel
size and version)
7 Vessel Sheath Viewing Lengthwise view glass with 179= 34 mm view
Window
8 Reserve port Cover port d =19 mm; septum e.g. for installation of 1- or 3-channel inoculation fittings
9 Reserve port Cover port d =19 mm; e.g. installation of Sarcova valve
10 Safety Valve | − Cover port d = 15 mm
Reserve Port − in ASME culture vessels: Cover port d = 19 mm
11 Supply Air Filter Cover port d = 19 mm, equipped with patented adapter for in-situ supply air filter sterilization
12 Reserve Port Cover port d = 19 mm, for level probe where necessary
13 Condensate Intake | Drain Only for stirrer shafts with double mechanical seal
40 Device Overview
3.5.2.2 Side Ports on the
Vessel
Upper port level (A)
Pos. Description
1..3 3 reserve ports, d = 25 mm, e.g. for intake valve APC 25 (accessory, optional)
Not pictured: Nozzle for burst disk ASME culture vessels
Lower port level (B)

Pos. Description
4 Port d = 12 mm, level, specially designed for Pt 100 temperature sensor
5 Port d = 25 mm, slanted, for pH or DO electrode or as a reserve port
7 Port d = 25 mm, level, e.g. for SVC 25 sampling valve
Vessel floor
1 Port d 32 mm for combined floor drain | sampling valve
Vessel wall
2 Port R 3/8“ connection of the double wall to the thermostat system
3.6 Agitator and The default top drive with brushless motor is detachably connected to the stirrer
Agitation Drive shaft with a flexible coupling. A single mechanical seal or double mechanical seal
pressure-superposed condensate receiver serves as the stirrer shaft seal.
Danger of burning skin upon contact!

− Avoid contact with the agitation drive when the agitator is in operation.
Wear safety gloves when working in the vicinity of the agitation drive.
Agitator with double mechanical seal (DGLRG)

The agitator and double mechanical seal can be operated either with 3-blade segment
agitators (CC variant) or with 6-blade disk agitators (MO variant).
The agitator with double mechanical seal is equipped with a sealing fluid system
that provides the necessary sliding film in the double mechanical seal. To do this,
condensate is obtained from steam by condensation. The condensate is pressurized
in the sealing liquid system (chapter “Operation”, section “double mechanical seal
(DGLRD)”.
If the double mechanical seal is operated without condensate pressurization,

this can damage the sliding rings.
t Check the fill level of the feed container before each process start.
t Sterilize and fill the feed container and the sliding ring seal.
t Pressurize after sterilization and fill the condensate feeder with pressurization
pressure.
Device Overview 41
Impeller
The following impellers are available as variants.
The impellers can be positioned freely on the agitator shaft. The impellers are secured
against slipping using clamping screws. Test the firm seating of the clamping screws
occasionally; they should be finger-tight.
Fig. 3-10: 3-Blade Segment Impeller Fig. 3-11: 6-Blade Disk Impeller
The stirring blades of the 3-blade segment impeller can be adjusted at an angle.
t Loosen the clamping screw on each stirring blade to do this.
t Adjust the angle of the stirring blade according to its requirement.
The default angle is 30 °.
Stirrer speeds
Culture vessel Maximum stirrer speeds BIOSTAT® Cplus
5L 1500 rpm
10 L 1500 rpm
15 L 1000 rpm
20 L 1000 rpm
30 L 600 rpm
3.7 Steam Distributor

The steam distributor is located to the left of the culture vessel. The steam valves for
components such as the floor drain valve seat bottom, APC's, and for the double
mechanical seal are located here.
The steam valves for sterilization of components are usually opened | closed using the
steam distributor.
Fig. 3-12: Steam Distributor
42 Device Overview
3.8 Manual Pressure
Control Valve
The manual pressure control valve is easily accessible in the supply unit in the exhaust
air segment downstream from the exhaust air filter.
You can use the pressure regulator valve to increase or decrease the pressure in the
culture vessel.
Fig. 3-13: Manual Pressure Control Valve
Device Overview 43
4. Transportation and Storage
4. Transportation and Storage
The device will be delivered by the customer service of
Sartorius Stedim Systems GmbH or by a transport company engaged by
Sartorius Stedim Systems GmbH.
4.1 Inspection Upon Acceptance

by the Recipient
4.1.1 Report and Document Upon acceptance of the device by the customer, the device must be inspected for
Transport Damage visible transport damage.
t Report transport damage immediately to the delivering office.
4.1.2 Check Completeness The delivery includes all required valves, connector elements, lines, hoses, and cables.
of the Delivery
t Check that the delivery is complete by comparing it against your order.
Components that do not correspond to the specifications of Sartorius Stedim Systems

GmbH may not be used.
4.1.3 Packaging The packaging used for transport and protection of the device consists primarily of
the following materials suitable for recycling:
− Corrugated cardboard
− Styrofoam
− Polyethylene film
− Pressed particle board
− Wood
Do not dispose of the packaging in the garbage.

Dispose of all packaging material in accordance with local regulations.
44 Transportation and Storage

4.1.4 Instructions for When moving the device, it is particularly important to do so in such a way as to
Transport within prevent damages by force or careless loading and unloading.
the Company
Danger of severe personal injury and property damage due to improper
transport!
− The device may only be moved by technical personnel.
− The load capacity of the lifting system (forklift) must be at least the weight of the
device. You can find weight specifications in the appendix [¨ Section “10.1
Specifications”].
− Wear protective work clothing, safety boots, safety gloves, and a hard hat during
these tasks.
− The device may only be transported with the transport locks in place. To install the
transport locks, contact Sartorius Stedim Service if necessary.
− Transport locks may only be removed at the place of installation.
− Lift the device only at suitable points with lifting accessories.
− Always lift the device slowly and carefully, to ensure stability and safety.
− Secure the device from falling during internal transport.
− During transport of the device, ensure that no personnel is in the path.
− These devices require two or more people to transport and set up.
− Wear protective work clothing and safety boots during these tasks.
− Lift the device only at suitable points.
− Always lift the device slowly and carefully, to ensure stability and safety.
− Secure the device from falling during internal transport.
− Engage the locking mechanisms on the device's castors after transport.
Protect the device during transport against:

− Moisture
− Impact
− Falling
− Damage.
Loading | Unloading
When loading and unloading, pay attention to the following aspects:

− Do not unload the device outdoors during rain or snow.
− If necessary, cover the device with plastic sheeting.
− Do not leave the device outdoors.
− Use only suitable, clean, undamaged load lifting accessories.
Transportation and Storage 45

4.2 Intermediate Storage If the device is not set up immediately after delivery, or will not be used temporarily,
the following conditions of storage must be noted:
Only store the device in dry buildings and do not leave the device outdoors.
In case of improper storage, no liability will be assumed for resulting damage.
46 Transportation and Storage

5. Setup, Assembly, and Initial
Commissioning 5. Setup, Assembly, and Initial
Commissioning
5.1 Setup | Assembly
5.1.1 Installation Location The guide for setup of the device is the setup drawing.
The setup of the device is to take place according to contractual conditions,
− by Sartorius Stedim Service,
− by Sartorius authorized specialist personnel,
The proper setup of the device is critical for its safe operation.
− Observe the guidelines for building and laboratory equipment.
− Observe the laboratory and process-related safety rules and guidelines on setting
up your workplace and securing it from access by non-authorized persons.
− Ensure that only authorized persons have access to the device.
− Follow the instructions in the following sections.
5.1.2 Ambient Conditions The device may only be operated under the following ambient conditions:
Criterion Ambient Conditions
Installation Location Conventional laboratory rooms, max. 2,000 m
above sea level
Ambient temperatures in the 5 – 40 °C
operating temperature range
Relative humidity <80 % for temperatures up to 31 °C (87.8°F),
decreasing linearly < 50 % at 40 °C (104°F).
Contamination Pollution degree 2 (normally only non-
conductive pollution occurs. Occasionally,
however, temporary conductivity caused by
condensation must be expected.)
Acoustic Emission max. sound pressure level < 80 db (A)
5.1.3 Acclimatization Condensation from humidity can form on the surfaces of a cold device when it is
brought into a substantially warmer area. You should therefore let a device that has
been disconnected from its power source acclimatize for approximately 2 hours
before reconnecting it to the power.
Setup, Assembly, and Initial Commissioning 47

5.1.4 Workplace Before setting up
Requirements
The working area at the device must be capable of being blocked off from
generally accessible areas.
Ensure that only authorized persons have access to the bioreactor.
Danger of injury if access to emergency shutdown equipment and shut-off

devices is blocked!
Equipment for emergency shutdown and shut-off devices, e.g., for the power supply,
water or gas feed, as well as the particular equipment connections, must be kept clear
and easily accessible.
t Follow the construction guidelines required to ensure that the device stands stably.
t Ensure that the foundation is dimensioned sufficiently for the weight of the device
and the process media in use.
t Ensure that the foundation is level.
t Ensure that the setup surface and room height are dimensioned in such a way that
the device is easily accessible for in-process operation, maintenance, and service
work. The space requirements also depend on the peripheral devices to be
connected.
5.1.5 Supply Facilities
5.1.5.1 Connecting Power/ The connections for energy and supply systems must be prepared before installation
Energy Supply Lines of the device in the work area, easily accessible, correctly installed, set in accordance
with the device's specifications, and ready to operate.
The connections for the supply media are located at the back of the supply unit.
Before connecting the energy supply lines to the bioreactor, the laboratory lines
should be flushed or allowed to discharge. If necessary, the suitable prefilters should
be installed.
Risk of injury from energy that is unexpectedly released, such as electric shocks!
Energy supply lines may be incorrectly dimensioned and not protected against
impermissible fluctuations and faults.
Safety equipment must be available and fully functional:
− Ground fault circuit interrupters (residual current protection) for mains
connections
− Fittings for shutting off water, compressed air and gas supplies.
Observe the energy specifications on the type plates [¨ see Section “5.1.5.2 Type
Plates”].
48 Setup, Assembly, and Initial Commissioning

Power

Causes severe injuries and can kill you.
Have the electrical connections installed by an electrician in compliance with VDE
and local guidelines, particularly safety measures.
The safety mechanisms required by safety guidelines and standards as used for
building installations must be present and functional.
For the protection of individuals, the electrical house connection must be protected
with a universal-current-sensitive ground fault circuit interrupter 3 + 30 mA.
t Make sure that network connections are secured against unacceptable voltage
fluctuations and surges.
t Ensure that the power connection is equipped with a ground fault circuit
interrupter.
t Ensure that the power connections are equipped with a power disconnecting
means on site.
t Ensure that there is good access to the power disconnecting means.
Supply Facilities
t Ensure that the intakes for cooling water, steam, compressed air, and gases
correspond to the specifications for the device [¨ see Section “10.1 Specifications”
and setup drawings and P&I diagrams in the “Overall documentation” folder),
without impermissible pressure variations.
t Ensure that the intakes are equipped with suitable valves for blocking and
emergency shutoff.
5.1.5.2 Type Plates The information on the correct supply voltage can be found on the manufacturer's ID
label of the supply unit. The manufacturer's ID label can be found at the rear of the
control/supply unit.
Fig. 5-1: Manufacturer's ID label control/ Fig. 5-2: Manufacturer's ID label control/
supply unit / version 208 V supply unit / version 400 V
Supply Facilities
t Ensure that the intakes for cooling water, steam, compressed air, and gases
correspond to the specifications for the device (for setup drawings and P&I
diagrams see the Technical Data Sheet for the device in the “Overall
documentation” folder), without impermissible pressure variations.
t Ensure that the intakes are equipped with suitable valves for blocking and
emergency shutoff.

5.1.5.3 Energy Supply Lines
Danger of functional problems and operational malfunctions or defects through
Specifications
contaminated cooling water or contaminated air and/or gases!
All media must be free of debris, corrosion residue, etc..
Demineralized water (e-water, deionized water), distilled water, pure water, salt water
or mineral water (especially if carbonated) are not suitable as cooling water.
Particle-containing air or gases are not suitable for aeration and pneumatic control.
Wet steam, steam with residual corrosion and using corrosion inhibitors whose
suitability has not been confirmed is unsuitable for temperature control and
sterilization.
Components of the coolant, such as antifreeze, must not damage the fittings,
particularly seals on pumps and valves.
At temperatures <4 ° lines and fittings can freeze. Condensation and condensate
must be able to run off without affecting or damaging equipment in the work
environment.
5.1.6 Disposal Equipment

Danger of infection from biologically contaminated exhaust air or waste water!
If exhaust filters are defective, valves in the intake or exhaust air paths are leaking, or
if there is improper operation of valves, biologically contaminated exhaust air may be
released. During manual sterilization of floor drain and sampling valves,
contaminated condensate may enter the drain.
− Follow the safety regulations relevant for the process.
− Set up the workplace to accommodate the requirements of the process.
− Set up and connect the appropriate equipment for collection and treatment of
contaminated exhaust air or waste water.
Containers or catch systems for biologically contaminated drain water and

condensate must have sufficient capacity and should be capable of being replaced
and if necessary separately sterilized.
− You can conduct the exhaust air from the vessel into the room or ambient air
through the sterile air filter. If you use laboratory facilities for the treatment of
contaminated air, these gas streams must be able to accommodate up to 50 l / min.
− Open laboratory drains for water and condensate must be designed for a flow of
about 1 l / min. The connection line must allow free flow without congestion
(water bags).
t Ensure that disposal systems are designed in accordance with the applicable legal
regulations and technical specifications.

5.1.6.1 Connections and Energy Supply Lines
Connected Item Electrical Characteristics Dimensions1) Special Notes

Compressed air 2) Feed 10 - 50 l/min; Quick Connect Coupling Air must be dry and free of dust and oil, Class 2
Pressure 4 ... 6 bar (g) Da=6 mm (ISO 8573-1)
Oxygen 2), Pressure 4 ... 6 bar (g) Quick connect coupling Gases must be dry and free of dust and oil
Nitrogen 2), CO22) Da=6 mm
Cooling water Inlet 3... 10 l/min; for hose 12+1/ Requirements:
Pressure 2 ... 4 bar (g) 15+1,5 mm 1) − particle-free tap water (if needed connect a 50 μm
prefilter);
− recommended hardness 12°dH
[¨ “Water Hardness Conversion Table”]
No demineralized or distilled water, e-water or
deionized water, salt water or carbonated water
When using ice water | cooling thermostats, avoid
the formation of ice; observe condensation and
condensate!
Water Return Connection to cooling for hose 12+1| Pressure difference Dp between flow and return at
circuit 15+1,5 mm 1) least 2 bar (g)
or drain 10 l/min
Pressure 2) Inlet 5... 15 kg/h; Clean steam, i.e. dry and free of particles,
(for heating) Pressure 3 bar (g); without corrosion-inhibiting agents or corrosive
ingredients
Pure steam Feed 3 kg/h; for tube 10+1 mm 1) Clean steam, i.e. dry and free of particles;
(for sterilization Pressure 2 bar (g); or hose nozzle if needed connect a 50 μm prefilter
of fittings)
Exhaust 2) in ambient air or laboratory Observe the requirements of the process-relevant
exhaust systems, 0 ... 50 l/ safety guidelines!
min Waste water temperature up to 95 °C
Condensate drain Open drain 1 l/min for tube 15+1,5 mm 1) Install drain that continually descends into a
(contaminated) laboratory drain to prevent backpressure.
Observe the requirements of the process-relevant
safety guidelines!
Mains connection 400 V / 50 Hz or 400 V: CEE plug, 5-pin Provision for the electrical house connection:
208 V / 60 Hz 208 V: NEMA plug, type Protection for personnel protection with universal-
L21-20P, 5-pin current-sensitive ground fault circuit interrupter
3 + 30 mA
1) When connecting via cutting ring screw connection in 120 V versions with NPT connection kit
2) Depending on the version
Water Hardness Conversion Table

Alkaline Alkaline German CaCO3 English French
earth ions earth ions hardness hardness hardness
[mmol/l] [mval/l] [°d] [ppm] [°e] [°f]
1 mmol/l alkaline earth ions 1.00 2.00 5.50 100.00 7.02 10.00
1 mval/l alkaline earth ions 0.50 1.00 2.80 50.00 3.51 5
1° German hardness [°d] 0.18 0.357 1.00 17.80 1.25 1.78
1 ppm CaCO3 0.01 0.020 0.056 1.00 0.0702 0.10
1° English hardness [°e] 0.14 0.285 0.798 14.30 1.00 1.43
1° French hardness [°f] 0.10 0.200 0.560 10.00 0.702 1.00

5.1.7 Setup Examples BIOSTAT® Cplus, bench-top system
Fig. 5-3: Dimensions of a BIOSTAT® Cplus
BIOSTAT® Cplus, bench-top system
Fig. 5-4: Dimensions of a BIOSTAT® Cplus with 5 L culture vessel

5.2 Starting Up
Running the system dry can cause damages to the thermostatic pump!
Temperature Control
Do not switch on the bioreactor until the temperature control circulation has been
Circulation
filled.
For initial startup of the bioreactor or after refitting and/or maintenance work,
the temperature control circulation must be filled with water.
Prior to filling, check that all fittings are tightened firmly. After filling, check for any
visible leaks. If there are, do not start up operation of the temperature control system,
but eliminate the cause of the leak.
2
Filling the temperature control circulation
1. Open the ball cock “cooling water supply” (1) and ball cock “cooling water
overflow” (2).
2. Add water until filling sounds are no longer audible. Water should come out of the
1 laboratory drain without bubbles.
Close the ball cock “cooling water overflow” (2).
The temperature control circulation has the pressure of the cooling water supply
(3 bar (g)).
3. Switch on the control unit. Check the temperature setpoint. To prevent
unnecessary heating at this time, set a setpoint in the range of ambient
temperature (about 20 ° C). Setup information for temperature control can be
found in [¨ Part B: Digital Measurement and Control Unit].
4. Let the thermostatic pump run for some time. Observe the pressure gage (4) on the
expansion tank (3). If a pressure drop occurs, bubble noises are audible or bubbles
appear in the water outlet, repeat steps 1 and 2.
4 5. Close the ball cock “cooling water supply” (1).
6. Open the ball cock “cooling water overflow” (2) again a little and observe the
pressure gage (3). Once it shows a pressure of 0.5 bar (g), close the ball cock
3 “cooling water supply” (1).
The thermostat cycle is ready for operation.

2
5.3 Initial Startup The initial commissioning of the device may only be carried out by Sartorius Stedim
Service or Sartorius-authorized technicians.

6. Operation
6. Operation
6.1 Safety Instructions Read the operating manual carefully before carrying out processes on the
device.
This is especially important for the safety instructions [¨ see Chapter “2. Safety
Instructions”].
6.2 Switching the Device Requirement

On | Off The system must be properly installed and connected in accordance with the
specifications. You must also have gained familiarity with the safety instructions in
[¨ Chapter “2. Safety Instructions”].
The switch-disconnector is the physical “mains in” on | off switch for the power
supply and is located on the left side of the control unit's housing. The switch-
disconnector is simultaneously the main switch used to turn the units on and off.
t Ensure that all required supply energies are connected to the device.
Switching on
1
t Turn the device on at the switch-disconnector (1).
Switching off
t Turn the device off at the switch-disconnector (1).
6.3 Triggering an In emergency or if a fault occurs, it is necessary to turn the device off immediately.
Emergency Stop
t Turn the switch-disconnector (1) to the zero setting to turn the device off.
The device can be turned back on after the emergency or fault is corrected (see
section “Turning the control unit on and off”).
6.4 Sensors The installation of sensors and calibration of sensors are described in the chapter
“7. Cleaning and Maintenance”.
54 Operation
6.5 Manual Pressure Control Valve
The manual pressure control valve is installed in the exhaust air line, the valve allows
you to raise or lower the internal pressure of the culture vessel.
Pos. Description
1 Hand wheel
2 Tube connector for exhaust
1 Pressure increase
t Turn the hand wheel clockwise to increase the pressure.
2 Pressure decrease
t Turn the hand wheel counterclockwise to decrease the pressure.
Fig. 6-1: Manual pressure control valve
Set the desired pressure on the pressure control valve; to do this, check the culture
vessel internal pressure on the gage, or in the DCU system if a pressure sensor is
installed, and correct it as needed by turning the handwheel.
Operation 55
6.6 Inserting Tubes into The peristaltic pump units are located on the control unit and convey the correction
Peristaltic Pumps media and nutrient media through hoses into the vessel.
Danger of crushing limbs by being pulled into the rotation pump!

Allow only qualified personnel to work on the device.
Disconnect the device from power while inserting hoses into the pump.
1. Open the flap. Press the hose clamp “inlet” and insert the hose.
2. Thread the hose into the rotor and turn the rotor clockwise by hand until the hose
is fitted in place.
3. Insert the hose into the outlet clamp and close the cover.
4. Check the contact pressure of the rollers in the pump head; they should clamp off
the hoses completely.
If the hose is pressed too tightly, it may wear out prematurely. Additionally, the
medium may not be allowed to backflow during breaks in pumping.
5. If necessary, correct the contact pressure at the adjustment screws.
In order to prolong the lifetime of the hose, you may occasionally stop the pump and
move the hose a bit. That way, the stress from flexion loading can be distributed over
longer hose sections.
For setting and operating the pumps, also observe the [¨ Pump manufacturer's
Fig. 6-2: Pump head of the hose pump: instructions].
– above: Inserting the hoses
– below: Setting the contact pressure
Pre-adjusting the Corrective Solution Supply
The tubes have different empty volumes depending on the their dimensions and
lengths.
The pumps must be temporarily activated to fill the hoses with corrective solution.
If the empty volume is not compensated for, the proportioning | feed volumes cannot
be calculated correctly.
1. Open the operating menu for each affected pump at the operating terminal and
chose the operating mode “Mode: on” [¨ Part B: Digital Measurement and Control
Unit].
2. If the transfer hose is filled to the inlet of the vessel, switch the pumps back to
“auto”. At this point, the corresponding regulator can activate the pump in order
to introduce acid, alkaline, antifoam agent or substrate.
If you do not need the corrective solution and would like to prevent inadvertent
introduction, you can select the operating mode “Mode: off” for the pump.
56 Operation
6.7 Internal Spinfilter The spinfilter is required to remove culture medium from the culture vessel during
continuous cell culture processes. The cells or the carrier material remain in the
culture vessel.
Depending on their use, spin filters are available with different mesh sizes. The mesh
size determines which components pass through the filter and which are retained in
the culture vessel.
The spinfilter is installed on the stirrer shaft. The cell-free culture medium is
harvested via a harvesting pipe spinfilter which is mounted in a 19 mm cover nozzle
with an inoculation fitting.
6.7.1 Design and Function Spinfilter
Pos. Description
1 Closing ring
2 Mesh
3 O-ring
4 Fixing screw
5 Spinfilter hub
Stirrer shaft and cover plate not pictured
Operation 57
Harvesting pipe spinfilter
2
3
Fig. 6-3: Harvesting pipe spinfilter design
Pos. Description
1 Septum holder
2 Inoculation nozzle
3 Septum
4 O-ring
5 O-ring
6 Harvesting pipe
Assembly position of the spinfilter and the harvesting pipe spinfilter

The spinfilter is mounted on the stirrer shaft so that the top edge is above the culture
medium. The culture medium may not flow over the edge into the inner space of the
spinfilter.
Where necessary, determine the respective maximum and minimum filling levels at
settings for maximum stirrer speed and maximum aeration.
Note that the filling volume can vary due to aeration, stirring and, where applicable,
additional equipment.
58 Operation
6.7.2 Installation and t Ensure that the spinfilter and the harvesting pipe spinfilter are installed.
Connection
Installing the septum
t Place a new inoculation membrane in the inoculation nozzle [¨ Fig. 6-3].
t Screw in a dummy plug.
The dummy plug fastens the inoculation membrane during vessel sterilization.
Preparing the inoculation fittings

t Stick the silicone hose in the harvesting bottle/bag.
t Cut the hose to the required length and connect it to the reservoir.
You can also use a sealable tube that is closed at one end.
t Screw the sterile sleeve onto the needle.
This will keep the inoculation needle sterile until you need the innoculation kit for
piercing and transfer of the seed culture or other media.
t Simultaneously, sterilize the inoculation fittings and the hose.
Connecting the inoculation fittings

t Unscrew the dummy plug from the inoculation nozzle.
t Singe the membrane briefly or spray it with a suitable disinfectant.
t Remove the sterile sleeve from the inoculation fitting.
t Singe the inoculation needle briefly and insert the needle vertically through the
membrane under flame protection.
t Screw the inoculation fitting into the inoculation nozzle using the cap nuts.
t Transfer the cell-free culture medium into the harvesting vessel.
Operation 59
6.8 Intake Units
6.8.1 Inoculation Port APC 19
6.8.1.1 Design and Function The APC 19 is a resterilizable inoculation port for installation into a cover port
d 19 mm.
The APC 19 consists of a:

− Resterilizable intake valve APC 19
− Steam valve on the steam distributor
− Condensate valve
− Intake valve with hose connector
− TC clamps and seals.
The connection to steam and condensate is carried out using pressure-tight, stainless
1 steel clad flexible hoses.
2
Pos. Description
1 APC 19
2 Intake valve
3 Condensate valve
3
Opening
t Turning the handwheel clockwise opens the APC 19
Close
t Turning the hand wheel counterclockwise closes the APC 19
6.8.1.2 Installation and Ensure that the APC 19 is correctly installed.

Connection t Close all manual valves.
t Connect the intake valve to a reservoir e.g. with a
− silicone hose,
− sealable hose with ventilation filter, e.g. Midisart® 2000, for later welding of
disposable bags.
t Autoclave the intake valve with installation parts.
Danger of burns!
Handle autoclaved, still-hot equipment carefully (e.g. during transport from the
autoclave to the workplace). Wear safety gloves.
t Install the autoclaved transfer valve back into the APC 19 intake group.
t Ensure that the hand valves are easily accessible.
t Check for firm seating of the TC connections and the pressure nuts on the
installation port.
60 Operation
Sterilization
Ensure that the APC 19 is correctly installed and that the intake valve and APC 19 are
closed.
Danger of burns!
Components become hot due to steam or hot condensate. Wear safety gloves.
Take measures to ensure that unauthorized persons do not tamper with the
components.
t Open the steam and condensate valve.
The steam intake should be open for about 20 min.

t Close the steam and condensate valve.
t Let the valve group cool before transferring media.
Transferring media from the reservoir

The media are transferred into the culture vessel using a peristaltic pump.
Ensure that the hose used is able to withstand the maximum operating pressure.
t Insert the hose according to [¨ Section “6.6 Inserting Tubes into Peristaltic
Pumps”].
Ensure that the steam and condensate valve is closed.

t Open the intake valve and the APC 19.
t Activate | deactivate the connected pump.
Emptying the transfer line

The transfer line should only be emptied at the end of the process.
t Ensure that the APC 19 and intake valve are open.
t Place the hose into the peristaltic pump in such a way that the medium is
conveyed back into the reservoir.
t Start the peristaltic pump and stop it when the hose is empty.
t Close the APC 19 and intake valve.
Operation 61
6.8.2 Intake and Sampling
Unit APC 25
6.8.2.1 Design and Function The APC 25 is an in-situ sterilizable intake port for installation into a side port
d 25 mm.
The APC 25 consists of a:

− Resterilizable intake valve APC 25
− Steam valve on the steam distributor
− Condensate valve
− Intake valve with hose connector
3 − TC clamps and seals.
1
Pos. Description
1 Intake valve
2 Condensate valve
3 APC 25
2
The connection to steam and condensate is carried out using pressure-tight, stainless
steel clad flexible hoses.
Opening
t Lift the locking lever (1).
t Press the valve lever (2) to the nozzle until the locking lever engages (latching
position b).
Close
t Lift the locking lever (1).
t Pull the valve lever (2) away from the nozzle until the locking lever engages
(latching position a).
62 Operation
6.8.2.2 Installation and Ensure that the APC 25 is correctly installed.
Connection t Close all manual valves.
t Close the intake valve on the reservoir e.g. with a
− silicone hose,
− sealable hose with ventilation filter, e.g. Midisart® 2000, for later welding of
disposable bags.
t Autoclave the intake valve with installation parts.
Danger of burns!
Handle autoclaved, still-hot equipment carefully (e.g. during transport from the
autoclave to the workplace). Wear safety gloves.
t Install the autoclaved transfer valve back into the APC 25 intake group.
t Ensure that the hand valves are easily accessible.
t Check for firm seating of the TC connections and the pressure nuts on the
attachment nozzle.
Sterilization
Ensure that the APC 25 is correctly installed and that the intake valve and APC 25 are
closed.
Danger of burns!
Components become hot due to steam or hot condensate. Wear safety gloves.
components.
t Open the steam and condensate valve.

t Close the steam and condensate valve.
t Let the valve group cool before transferring media.

Ensure that the hose used is able to withstand the maximum operating
pressure.
Pumps”].
Ensure that the steam and condensate valve is closed.
t Open the intake valve and the APC 25.
t Activate | deactivate the connected pump.

t Ensure that the APC 25 and intake valve is open.
t Close the APC 25 and intake valve.
Operation 63
6.8.3 STT Quick Connector The STT quick connect coupling can be used to produce fast and secure sterile hose
connections. This way, lines and vessels can be connected aseptically for inoculations,
supplying corrective solutions or transferring extracted culture media.
The coupling part of the STT quick connect coupling is normally connected to the
vessel line, the plug to the transfer line of the reservoir vessel or the harvest
container.
6.8.3.1 Equipment and Item no. Description

Specifications
8809240 STT quick connect coupling half (female)
− Autoclavable quick connect coupling half made of stainless steel
− for sterile connection of hoses with an internal diameter of
3.2 ... 5.0 mm
− Equipment supplied: Female coupling half, 1 dummy plug
8809208 STT quick connect coupling half (male)

3.2 ... 5.0 mm
− Equipment supplied: Quick connect coupling, male half, 1 cap
8809410 STT quick connect coupling half (female)

1.6 ... 2.0 mm
8809402 STT quick connect coupling half (male)

1.6 ... 2.0 mm
64 Operation
6.8.3.2 Assembly
Prepare the reservoir vessels or harvesting bottles and the connection to the vessel in
such a way that the STT plug-in nipple is positioned at the line to exernal vessels and
the STT coupling at the connection to the vessel. Connect the STT components prior
to sterilization.
Assembly and Connection of the STT coupling part (7):

1. Place the slit membrane (2) into the membrane holder (1).
Fig. 6-4: Coupling part of the STT coupling
2. Screw the adapter (3) onto the membrane holder (1).
3. Attach a piece of silicone tubing (4) to the coupling part and the inoculation
fitting to connect it to the vessel.
4. Insert the dummy plug (5). This maintains sterility after autoclaving until the plug
is connected.
5. Autoclave STT coupling part with the transfer line and the inoculation fitting.
Connect STT plug to the supply vessel:

1. Cut a sufficiently long piece of silicon hose (10) to use as a transfer line and attach
it to the plug-in nipple (8) as well as to the harvesting tube of the vessel.
2. Close plug-in nipple (8) with the sterile cap (9). The cap protects the plug-in nipple
from contamination until the seed material | media can be transferred.
Fig. 6-5: Male part of the STT coupling
Connecting the STT quick connect coupling

1. Remove dummy plugs from the sealing coupling (7) and the sterile cap from the
plug-in nipple (8).
2. Insert plug-in nipple in the coupling and push it through the slit membrane.
Press the pieces together and turn them until the cylinder pin (6) locks.
Fig. 6-6: Connect STT coupling parts
6.8.3.3 Transferring a Medium 1. To transfer the media, lift reservoir vessel or connect the hose to a peristaltic
pump.
2. A peristaltic pump can be used to extract a sample and transfer the medium into
a harvest container.
3. Pull the plug-in nipple out of the coupling part if you wish to connect an
additional line. Connect the next inlet or close the coupling part with the dummy
plug, until you need to use the connection again.
6.8.4 Inoculation Fittings 1- and 3-channel inoculation fittings

and Septa The inoculation fittings permit the vessel to be pierced during the process without
danger of contamination, in order to add inoculation cultures or correction media
and other special media, or to remove them using a harvesting pipe. You need a
19 mm port (septum) in the cover plate for the connection.
Important information!
Operation 65
Septa (in the cover plate)
Using septa, inoculation cultures and other media can be added to the process in the
vessel or harvested without danger of contamination.
The media are transferred into the vessel using a syringe or an inoculation fitting.
During the next cultivation run, the membranes used must be replaced.
Installing the septum

t Insert a new inoculation membrane into the d 19 mm port.
t Screw in the membrane connection nozzle (2) and tighten it.
This fastens the inoculation membrane.
t Screw in a dummy plug (3).

t Sterilize the culture vessel.
The dummy plug fastens the inoculation membrane during vessel sterilization.
Fig. 6-7: Septum in the cover plate
Preparing the inoculation fittings

t Stick the silicone hose to the reservoir bottle or container for the inoculation
culture onto the hose nozzle (4).
t Cut the hose to the length required and connect it to the reservoir, or use a
weldable hose that is closed on one end.
t Screw the sterile sleeve (5) onto the inoculation needle. This will keep the
inoculation needle sterile until you need the innoculation fitting for piercing and
transfer of the seed culture or other media.
t Sterilize the inoculation fitting with the hose or with connected peripherals.
Connecting the inoculation fittings

t Screw the dummy plugs (6) out of the membrane connection nozzle.
t Singe the membrane briefly or spray it with a suitable disinfectant.
t Remove the sterile sleeve (5) from the inoculation fitting (1).
t Singe the inoculation needle briefly and insert the needle vertically through the
membrane under flame protection.
t Screw the inoculation fitting into the membrane holder (7) using the cap nuts (2).
t Transfer the inoculation culture or medium.
t Afterwards, you can leave the inoculation needle in the membrane holder (7) and
the hose clamped off.
Fig. 6-8: Inoculation fitting
66 Operation
6.8.5 SACOVA Valve
SACOVA valves must be installed before vessel sterilization.
You can use the SACOVA valve to transfer inoculation cultures, correction media,
and nutrient media into the vessel without contamination, and without piercing.
SACOVA valves can be installed in ports in the cover plate (d 19 mm) or in the side
nozzle (d 25 mm). Variants with 1-channel intake or 3-channel intake are available.
Preparing for sterilization in autoclaves

Put the SACOVA valve into the closed position.
Turn the knurled nut (1) up and pull the valve tappet (2) up (position “A”).
t Close the SACOVA valve on the reservoir e.g. with a

− silicone hose,
− weldable hose with ventilation filter, e.g. Midisart® 2000, for later welding of
feed containers/bags.
Fig. 6-9: 3-channel SACOVA valve
t Autoclave the SACOVA valve in the closed position with peripherals connected.
Installation into the vessel:

t Screw the autoclaved SACOVA valve into a free cover nozzle d 19 mm).
t Screw the SACOVA valve in firmly with the pressure nut (2).
t Leave the valve tappet in the closed position “A”.
t Sterilize the culture vessel.

Pumps”].
t Open the SACOVA valve by turning the knurled nut (1) in the clockwise direction
until it stops.
y The SACOVA valve is now in the “B” position (open).
t Activate the transfer of the medium.
If you are using peristaltic pumps or other positive displacement pumps to transfer
the medium:
Hoses may break and media may escape in an uncontrolled manner!

If you turn on the pump or the regulation circuit in question with the SACOVA valve
in the “closed” (”A”) position, an impermissible overpressure may result.
Operation 67
t Ensure that the SACOVA valve is open.
t Close the SACOVA valve.
6.8.6 Correction The correction media bottles can be used for acids, bases, anti-foam agents, and
Media Bottles nutrient solutions. They are delivered ready to use and sterilized with the correction
medium or nutrient solution in autoclaves.
Danger of injury from shattered glass and escaping media!

Damaged glass bottles can break in autoclaves or due to incorrect handling. The
media, for example acids or bases, can be released unintentionally. Defective hoses
can also release media.
Carefully clear away the broken glass and spilled media as quickly as possible.
Danger of chemical burns from handling acids or bases!

Escaping acids and bases can burn body parts or clothing.
Use protective clothing, safety gloves, and safety glasses.
Components made of unsuitable materials can be damaged!

Use only parts that are resistant to the media. Avoid the use of hydrochloric acid (HCl)
for regulation of pH. HCl can also attack stainless steel parts.
Handle glass bottles carefully:
Replace damaged bottles.
Regularly check the gaskets and transfer hoses for damage and replace them if
necessary.
Replace the ventilation filters before any sterilization in autoclaves.
68 Operation
Preparing the correction medium bottle
When carrying out lengthy or continuous processes, you should prepare multiple
bottles in order to have sufficient sterile solution, or use disposal bag systems.
t Place the PTFE tube (7) onto a hose nozzle on the bottom.
Shorten it until it reaches to about 1 – 2 mm above the floor of the bottle.
t Fill the bottle (1) with anti-foam solution, acid, base, or substrate.
Place the silicone gasket (2) and the head piece (3) onto the edge of the glass and
close the bottle with the screw cap (4).
The bottle can be connected to the following components for transfer of media:
− Inoculation fittings (inoculation ports)
− SACOVA valve
− 4-valve intake group
t Place a piece of silicone hose (6) onto the hose nozzle to which the PTFE pipe (7) is
attached.
The transfer hose must be long enough that you can insert it into the associated
peristaltic pump.
t Place the sterile filter (5) and silicone hose onto the remaining hose nozzle on the
bottle.
The sterile filter must be replaced before any sterilization in the autoclave.
t Connect the free end of the transfer hose to an inoculation fitting, a SACOVA
valve or a 4-valve intake group.
t Secure all hoses with hose clamps.
t Autoclave the correction medium bottle including the intake valve.
t Clean the bottle after the process finishes.
Fig. 6-10: Bottle for sampling
Operation 69
6.9 SVC 25 Sampling Valve The sampling valve SVC 25 is used for precise sampling from fermenter vessels.
The valve fits into a 25 mm port and can be sterilized in-situ with steam.
Equipment features include:

− Sampling valve
− Standard
− Containment sampling
6.9.1 Sampling Valve Standard, The sampling system consists of the sampling valve, a steam line with steam valve,
Design | Function a sterile sleeve, and TC clamps and gaskets.
Pos. Description
1 Sampling valve
2 Steam valve
2 3 Drain arc
4 Sterile sleeve
Opening
1 t Turning the hand wheel clockwise opens the sampling valve.
3 Close
t Turning the hand wheel counterclockwise closes the sampling valve.
Fig. 6-11: Sampling valve with sterile sleeve
6.9.1.1 Installation and

Only install the SVC 25 in a horizontal 25 mm port of the culture vessel.
Connection
Ensure that the sampling valve is correctly installed.

t Close the sampling valve.
t Plug the sterile sleeve into the drain pipe.
t Place a metal bucket under the outlet of the sterile sleeve.
70 Operation
Sterilization
Danger of burns on valves!

Use safety gloves when manipulating the valves.
If sterilization is interrupted, wait until a safe state has been achieved (cooled to
ambient temperature, pressure released) and then continue with the work.
Ensure that the sampling system is correctly installed.

t Carefully open the steam valve on the steam distributor and the steam valve (2).
Only a little steam should escape from the outlet of the sterile sleeve (4).
Adjust the regulation if necessary.

t Close the steam valve on the steam distributor and the steam valve (2).
t Allow the sampling valve to cool.
Connect a silicone hose (length from floor to bucket + 20 cm) to the hose connector
of the sterile sleeve. This will reduce the noise during sterilization.
Taking a sample (sampling)

t Remove the sterile sleeve (4).
t Place a container under the outlet of the sampling valve.
t Open the sampling valve.
t Remove the desired quantity of liquid and close the sampling valve.
t Attach the sterile sleeve (4).
t Sterilize the sampling system.
Operation 71
6.9.2 Containment Sampling, The containment sampling system consists of the sampling valve, a steam line with
Design | Function steam valve and condensate line with condensate valve, a containment sampling
bottle and TC clamps and gaskets.
Pos. Description
2 1 Sampling valve
2 Steam valve
3 Drain arc
4 Sampling bottle with valve
5 Condensate valve
1
Opening
t Turning the hand wheel clockwise opens the sampling valve.
3
Close
5 t Turning the hand wheel counterclockwise closes the sampling valve.
Fig. 6-12: Sampling valve with sterile sleeve
Preparing the sampling bottle

For contamination-proof sampling, you can connect the bottle to the vessel using the
sampling valve.
t Ensure that the sampling bottle is clean.

t Check the cover seal for damage and replace it if necessary.
3
t Screw the cover onto the bottle and ensure that the cover seal is seated properly.
The sterile filter (1) must be replaced before any sterilization in the autoclave.
t Ensure that the connection hose (2) from the sterile filter to the bottle is not
1
kinked and the valve (3) is closed.
2
The sterile filter will ensure that pressure compensation can take place in the bottle
during sterilization in the autoclave.
Fig. 6-13: Sampling bottle valve / filter
72 Operation
6.9.2.1 Installation and Ensure that the sampling valve is correctly installed.
Connection
t Close the sampling valve.
t Install the sampling bottle according to the instructions.
t Autoclave the sampling bottle.
t Connect the sampling bottle to the sampling valve.
t Connect the condensate line to the sampling bottle using the condensate valve.
Sterilization

The containment sample can be carried out together with the culture vessel or
separately later.
Sterilizing separately
Ensure that the sampling system is correctly installed.
t Open the valve on the steam distributor as well as the steam valve (2) and the
condensate valve (5).
t Close the steam valve (2) and the condensate valve (5) as well as the steam valve
on the steam distributor.
t Carefully open the valve on the sampling bottle to avoid the formation of a
vacuum.
t Allow the sampling valve to cool.
Taking a sample (sampling)

t Open the sampling valve (clockwise rotation).
t Remove the desired quantity of liquid and close the sampling valve.
t Connect a newly autoclaved sampling bottle to the sampling valve.
t Sterilize the sampling system.
Operation 73
6.10 Floor Drain Valve The floor drain valve is screwed or welded into the floor of the vessel in a centric
orientation. Complete emptying is guaranteed.
The floor drain valve is manually operated and is suitable for sampling or for draining
the culture vessel contents.
2
2b 2a
Fig. 6-14: Floor drain valve / functional elements
Pos. Description
1 Release line (Adapter and TC connections are not shown)
2 Sterile sleeve
2a Counterclockwise rotation (close)
2b Clockwise rotation (open)
3 Hand wheel
4 Floor drain valve
Preparing the floor drain valve

Ensure that the floor drain valve is correctly installed.
t Close the floor drain valve.
t Attach the sterile sleeve.
t Place a metal bucket under the outlet of the sterile sleeve to catch the emerging
steam/condensate.
Sterilizing the floor drain valve

Ensure that the floor drain valve is correctly installed.

t Close the floor drain valve.
t Attach the sterile sleeve.
t Place a metal bucket under the outlet of the sterile sleeve.
t Carefully open the steam valve on the steam distributor. Only a little steam should
emerge from the outlet of the sterile sleeve; readjust it if necessary.
t Leave the steam intake open for about 20 min.
t Close the steam valve.
t Allow the floor drain valve to cool.
74 Operation
Connect a silicone hose (length from floor to bucket + 20 cm) to the hose connector
of the sterile sleeve. This will reduce the noise during sterilization.
Sampling / harvesting
t Remove the sterile sleeve (2).
t Place / Connect a vessel at the outlet on the floor drain valve.
t Open the floor drain valve.
t Remove the desired quantity of liquid and close the floor drain valve.
t Replace the sterile sleeve (2).
t Sterilize the floor drain valve.
6.11 Dummy Plugs Unneeded cover penetrations and side ports must be sealed with dummy plugs.
t Insert the dummy plug (1) into the unneeded cover penetration (2) or side port (3).
t Hand tighten the screw connector (if installed in the cover) or the cap nut
(if installed in a side port).
Fig. 6-15: Dummy plugs and connections
Pos. Description
1 Dummy Plugs
2 Cover port
3 Side port
4 Cap nut
5 O-ring
Operation 75
6.12 Sterilization
6.12.1 Safety Instructions Danger of injury in the vicinity of the bioreactor!

The vessel and in-situ sterilized components and lines are heated to the sterilization
temperature and are under pressure. Parts that are incorrectly installed or mani-
pulated, as well as steam and | or hot culture medium, can be explosively ejected.
Scratches or hairline cracks on glass containers (correction medium and sampling
bottles) can impair their pressure resistance to the point that operational safety can
no longer be ensured for sterilization. Handle the culture vessel very carefully.

If sterilization is interrupted, wait until the bioreactor has achieved a safe state
(cooled to ambient temperature, pressure released) and then continue with work on
the vessel.
Before each sterilization, check:

− The installation of parts and connections to the vessel.
− Required safety systems; burst disks and | or safety valves must be installed.
− Install the splinter protection sheath in 5 L culture vessels.
Restrict presence at the bioreactor to the necessary operator interventions.

bioreactor.
Block off the danger zone and attach suitable, easily visible warning signs.
Start (and stop) the sterilization only using the sterilization program of the control
unit (except by manually actuated valves such as sampling and intake valves or the
double mechanical seal, no user intervention is required).
76 Operation
6.12.2 Splinter Protection
Setup for 5-liter
Culture Vessels
Attach the splinter protection before each sterilization of the culture vessel.
1. Place the splinter protection sheath around the glass cylinder and adjust it so that
it rests evenly on the base section.
2. Hang the hooks (3) in the eyelets (2). You should be able to do this easily. If not,
re-adjust the sheath.
3. After sterilization, you can remove the splinter protection when the medium has
cooled to fermentation temperature.
Pos. Description
1 Splinter protection
2 Mounting eyelet
3 Hooks
4 Base section flange
5 Base section of the vessel
6.12.3 Adjusting Components t Check whether all components and accessory parts have been installed that you
will need for the process and that must be installed before sterilization.
t The pH sensor must be calibrated. If necessary, do this before filling the vessel.
t Close the floor drain valve and any ports and accesses that are still open.
t Close the sampling valve and | or the built-in intake valve.
t Install the splinter protection sheath in 5 L culture vessels [¨ see Section “6.12.2
Splinter Protection Setup for 5-liter Culture Vessels”].
Operation 77
6.12.4 Carrying Out Sterilization of the culture vessel takes place in multiple steps in a defined sequence
Sterilization and is started and stopped from the operator terminal.
The following assemblies are sterilized together with the culture vessel:
− Aeration segment
− Exhaust air segment
− Built-in assemblies such as sensors, agitator, etc.
− Intake valve APC (on vessel)
− Floor drain valve (on vessel)
− Sampling valve (on vessel)
The following components, if present, must be sterilized manually after sterilization

of the culture vessel:
− Intake valve APC (on intake side)
− Floor drain valve (exhaust side)
− Sampling valve (sampling side)
− Sealing liquid system of the double mechanical seal
6.12.5 Sterilizing the

If the double mechanical seal is operated without condensate pressurization,
Double
this can damage the sliding rings.
Mechanical Seal
Check the filling level of the sealing liquid container before each process start and
sterilize and fill the sealing liquid container and the sliding ring seal.
After sterilizing and filling, fill the fittings with pressurization pressure.
6.12.5.1 General Notes − You can sterilize the double mechanical seal in the stopping phase of the
sterilization of the vessel. This is not necessary during each sterilization.
− Check the level in the sealing liquid container regularly. Slight condensate
consumption is normal and escaping condensate is not an indication of a defect.
One filling of the condensate reservoir normally suffices for several processes.
− A filter in the condensate reservoir prevents dirt particles from entering the double
mechanical seal and damaging the contact faces. In general, you should use pure
steam for sterilization of double mechanical seal and the production of
condensate.
78 Operation
6.12.5.2 Sterilizing the Double Overview
Mechanical Seal
1 2 3 4
Fig. 6-16: Schematic design of the double mechanical seals with fittings
Pos. Description
1 Vent valve
2 Viewing window
3 Air supply valve
4 Air supply gage
5 Steam trap valve
6 Steam supply valve
t Close the air supply valve (3).
Operation 79
t The vent valve (1) must be closed.
t Open the steam supply valve (6) and steam trap valve (5) during the vessel
sterilization stopping phase.
y The orifice (TH-203) limits the steam pressure to about 1.5 bar (g).
t Steam for at least 20 minutes to achieve sufficient sterilization.
6 5 t Close the steam trap valve (5) during the vessel sterilization cooling phase.
t Connect the cooling water supply.

Remove the supply line from the exhaust cooler and connect it to the condensate
reservoir.
t Open the cooling water supply (7).
y Now steam can condense in the condensate reservoir and fill it with sterile
7 condensate.
t Fill the sealing liquid vessel to about 3/4 of the height of the view glass (2).
t Close the steam supply valve (3).
4 t Disconnect the cooling water supply again.
t Briefly open the valve to vent and close it again.
t Check the gage (4). The pressure should be approx. 1.4 bar (g).
80 Operation
t Slowly open the air supply valve (3).
6.12.5.3 Full Sterilization
To compensate for the vacuum in the vessel after sterilization, the culture vessel
must be aerated with 0.5 vvm (based on the max. working volume of the culture
vessel) at a preliminary pressure of 1.5 bar (g). Setup must take place at the
beginning of vessel sterilization.
t Fill the culture vessel with at least 50% of the maximum operating volume.
Note that liquid will vaporize during sterilization.

The loss due to vaporization can only be determined by observation.
If your system also has the empty sterilization function, steam (and thus condensate)
is also conducted into the culture vessel during full sterilization. The resulting fluid
loss / gain can only be determined empirically.
Ensure that all culture vessel openings are closed and attachments are correctly and
firmly attached.
t On the air rotameters (sparger & overlay), set an overall gas flow of 0.5 vvm based
on the max. operating volume of the culture vessel.
For gas paths with an installed mass flow controller, completely open the precision
adjustment valve on the rotameter.
1 t Lift the supply air filter adapter “sparger” (1) into the “sterilization” position.
t Close the ball cock on the cooling water supply to the exhaust cooler (1).
1
t Start the full sterilization sequence at the control system.
Operation 81
6.12.5.4 Empty To compensate for the vacuum in the vessel after sterilization, the culture vessel
Sterilization must be aerated with 0.5 vvm (based on the max. operating volume of the
culture vessel) at a preliminary pressure of 1.5 bar (g). Setup must take place
at the beginning of vessel sterilization.
Ensure that all culture vessel openings are closed and attachments are correctly and
firmly attached.
t Ensure that the steam distributor valve (1) is open.

1
t On the air rotameters (sparger & overlay), set an overall gas flow of 0.5 vvm based
on the max. operating volume of the culture vessel.
For gas paths with an installed mass flow controller, completely open the precision
adjustment valve on the rotameter.
2
t Connect the condensate line (1) to the outlet of the floor drain valve.
1 t Open the floor drain valve (2).
t Lift the supply air filter adapter “sparger” (1) into the “sterilization” position.
1
t Close the ball cock on the cooling water supply to the exhaust cooler (1).
t Start the empty sterilization sequence [¨ see Part B: Digital Measurement and
1 Control Unit] at the control unit.
t Close the floor drain valve after the sterilization sequence has ended.
82 Operation
6.13 Performing Processes
6.13.1 Sterile Test and Contamination by external germs generally leads to failure of the process. Causes for
Pressure-hold Test such contamination can include insufficient sterilization of the medium, damaged
gaskets or filters, or improper installation of vessel equipment.
Since not all noticeable changes are indications of an infection, if there is a suspicion
of contamination you should perform additional investigation.
t Before the process, carry out a sterile test or pressure-hold test.
All equipment and peripheral connections to the vessel must be connected and
the process conditions (e.g. temperature, aeration, etc.) must be configured.
6.13.1.1 Carrying Out t Allow the system to operate for about 12-24 hours and observe the pH value,
a Sterile Test the DO value, and turbidity in the nutrient medium in the culture vessel.
− Different pH values before and after sterilization can be due to chemical reactions
in the medium. If the pH value continually drifts during the sterile test, this may be
an indication of contamination.
− Changes in the DO value after aeration starts can be due to chemical reactions.
If the oxygen consumption increases linearly or exponentially during the sterile
test, this indicates contamination.
− Cloudiness of the medium can be due to chemical reactions or the agglomeration
of medium components and need not be the result of contamination.
If contamination is determined and the sterile test must be interrupted, proceed as

follows:
t Sterilize the culture vessel and its equipment and empty the vessel.
t Check that the installed parts are firmly seated.
t Check the gaskets. Clean dirty gaskets and replace damaged ones, for example if
pressure marks indicate damage.
t Check the intake and exhaust air filters and replace them if necessary.
t Repeat the sterile test.
If contaminations continue to occur, you can extend the sterilization time. Increase
the sterilization temperature only if the vessel equipment is suitable for temperatures
> 121 °C.
Each manipulation of the vessel, vessel equipment, and feed lines risks the
introduction of germs. To limit the cause of a contamination, you can take regular
samples and check for external germs:
− Before inoculation from the culture medium and the correction medium bottles
− After inoculation from the vessel and non-transferred residue of the inoculation
culture
− After sampling from the culture, from samples needed for other investigations.
Operation 83
6.13.1.2 Carrying Out the The pressure-hold test is used to check the device for leaks.
Pressure-hold Test
The device can be equipped with an automatic pressure-hold test.
The pressure-hold test is started from the control unit [¨ see Part B: Digital
Measurement and Control Unit].
Automatic pressure-hold test

The automatic pressure test requires the following equipment components to be
installed in the device:
− Pressure sensor
− Pressure control valve
− Software sequence for pressure-hold test
Pressure loss
If you detect a pressure loss, locate the leak and correct the problem. To do so,
see [¨ Chapter “8. Disruptions”].
6.13.2 Preparing the Check the following settings and connections and modify them if necessary for the
Bioreactor for process:
the Process
t Allow the culture vessel to cool to the specified operating temperature after
sterilization.
t Connect the quick-release couplings to the exhaust air cooler and open the
cooling water valve, if you have used the cooling water connections of the exhaust
air cooler to generate condensate for the double ring seal in the meantime.
t Calibrating the slope of the DO sensor [¨ see Section “7.7.7 Sensors”].
t Connect the separately sterilized correction medium bottles.
t Insert the hose into the peristaltic pump.
t Open the intake groups if needed.
t Configure the process parameters for fermentation in the control unit and turn on
the required controllers (instructions for setting the parameters can be found in
[¨ Part B: Digital Measurement and Control Unit]):
− Operating temperature
− Impeller speed
− pH value
− DO value
− Anti-foam regulation
− Level regulation
− Operating pressure
Agitator speed
Danger of vibrations and damage to vessel attachments from excessive speed.
84 Operation
6.13.3 Inoculating the t Transfer the inoculation culture into the culture vessel using a SACOVA valve,
Culture Vessel an inoculation fitting, or an APC.
Follow the instructions in the section for each intake group.
t Allow the inoculated fluid to run in under gravity or transfer it into the culture
vessel with a peristaltic pump.
Introducing media
The introduction of nutrient media and correction media such as acid, base, anti-
foam agent into the culture vessel can be done using a SACOVA valve, inoculation
fitting, or an APC.
Follow the instructions in the section for each intake group.

t Introduce media into the culture vessel using a peristaltic pump.
The speed limitation is pre-configured for each bioreactor.

Make sure not to exceed the maximum permissible impeller speed (see technical
information).
6.13.4 Completing the Process t Harvest or transfer the culture broth using the floor drain valve.
t Remove the inoculation fittings and close the ports with dummy plugs.
t Sterilize the culture vessel and attachments if necessary.
t Clean and | or maintain the entire system.
Operation 85
7. Cleaning and Maintenance
7. Cleaning and Maintenance
Incorrect cleaning and maintenance can lead to erroneous process results, causing
high production costs. Regular cleaning and maintenance is thus essential. Among
other factors, the operational safety and effective performance of fermentation also
depend on proper cleaning and maintenance.
The cleaning and maintenance intervals largely depend on the stress placed on the
culture vessels and equipment by aggressive components contained in the media
(e.g. acids and bases used to regulate pH) and the level of contamination from culture
and metabolic product residues attached to this equipment.
7.1 Safety Instructions Danger to life caused by electrical voltage!

Electrical switching elements are installed in the device. Contact with parts under
voltage represents a direct danger of death.
− Never open the unit. The unit may be opened only by authorized personnel of the
Sartorius Stedim Biotech Company.
− Work on the electrical equipment of the device may only be carried out by
Sartorius Stedim Service or authorized technicians.
− During maintenance and cleaning, turn the power supply off and secure it against
reactivation.
− Keep moisture away from parts under voltage, as it can lead to short circuits.
− Check the electrical equipment of the unit regularly for defects such as loose
connections or damage to the insulation.
− In case of defects, turn the power supply off immediately and have the defects
corrected by Sartorius Stedim Service or authorized technicians.
− Have the electrical components and stationary electrical equipment checked by an
electrician at least every 4 years.
Possible biohazard!
(depending on microorganisms or cells)
Follow relevant safety guidelines.
Sterilize the vessel with all attachments after completion of the process and harvest
of the culture medium.
Danger of limbs being pulled into the rotation pump and crushed.
− Allow only qualified and authorized personnel to work on the unit.
− Disconnect the unit from power when performing maintenance and cleaning tasks.
Danger of burns upon contact with hot surfaces.

− Avoid contact with hot surfaces like vessels, motor housings, and pipelines carrying
steam.
− Wear protective gloves when working with hot culture media.
86 Cleaning and Maintenance

Danger from projecting components!
− Ensure that danger points such as corners, edges, and projecting components are
covered.
Preliminary Steps
Always take the following preliminary steps when performing cleaning and
maintenance:
t Turn the device off at the main switch.
t Remove the power supply from the laboratory connection.
t Turn off all supply media in the lab (water and gas supply).
t Ensure that the connections and hoses have been depressurized.
t If necessary, remove the supply media lines from the device.
7.2 Assembly / Disassembly To disassemble of the cover plate, it is necessary to remove the motor from the cover
of the Motor plate. To do this, proceed as follows:
Risk of injury!
Take note of the motor’s and the cover plate's weight. Always use suitable lifting
equipment.
The cover plate's O-ring may stick. Loosen the cover carefully so as not to damage
the O-ring. Do not knock the internal components attached to the cover against the
vessel.
Disassembly
t Loosen the screws (4) affixing the guide sleeve of the motor to the stirrer shaft
coupling. Lift and lower the motor carefully.
Mounting the motor (after installation of the cover plate; [¨ see Section “7.4
Disassembly / Assembly of the Cover Plate”]).
t Place the motor (2) on the vessel with the guide sleeve on the stirrer shaft adapter
(1) so that the pin (3) locks into place.
t Stick the screws (4) into the stirrer shaft bearing sleeve bores through the guide
sleeve on the engine and tighten down the motor with them.
Cleaning and Maintenance 87

7.3 Cover Lifting System The cover lifting system allows you to lift the culture vessel cover out of the vessel
safely and with little effort and mount it safely back on the vessel after conversion
and maintenance work, such as on the stirrer shaft or other vessel equipment.
The cover lifting system is mechanical and is operated manually by a hand crank:
Danger of crushing by vessel cover!

Extremities may be crushed during assembly and disassembly.
Only hold the cover using the handles provided.
Two people working together should lift the cover.
Dismounting/mounting with lifting accessories:
Use only the lifting accessories provided to lift the cover.
Attach the lifting accessories only to the fastening points provided.
Risk of injury from improperly attached falling parts!

Only individuals working on the unit may be within your work area.
Block the work area off.
Individuals carrying out the work must wear safety shoes.
The cover lifting system requires two people to operate:

− 1 person to operate the lifting system,
− 1 person to guide and direct the cover when lifting and lowering so that it does
not run into housing parts or the vessel (valve assemblies on the boiler).
7.3.1 Lifting the Cover Plate

off of the Vessel
4
1 Motor
3
3 5

A holder locking pin (1) on the lifting arm stand locks the lifting arm in two positions:
− [Ä] shows the pin in its resting and maintenance position (locked),
− [Ã] is the position for raising and lowering the cover plate (unlocked).
Danger of injury; cover plate may fall if sleeve is unsecured!

Lightly crank the lifting system until the cable is taut and make sure that the sleeve is
mounted correctly on the stirrer shaft coupling.
t Pull locking pin (1) out into position [Ã].

t Pivot lift arm by lateral rotation (2) on the crankcase until the lifting system hangs
centrally over the cover plate.
t Engage the locking pin in position [Ä].
t Lower the sleeve of the lifting system onto the stirrer shaft coupling and engage.
Then carefully attach the sleeve with the two safety screws.
t Loosen cover screw connections with a suitable tool, e.g. a wrench.
t Carefully turn the crank of the lifting system to lift the cover plate.
The cover plate seal may stick to the vessel flange.

t In this case, carefully turn the cover sideways or shake it loose.
t Turn the crank further and lift the cover with stirrer shaft, sparger pipe and the
still-mounted vessel components.
The second person should guide the cover plate and prevent the cover from
running into the bioreactor frame, vessel attachments, the inner wall of the vessel
or the vessel flange.
t Once stirrer shaft and sparger are above the vessel flange, pull the locking pin out
of its engaged position [Ã] and pivot the lifting arm sideways.
t When the locking pin snaps back into position [Ä], the cover plate is in a position
it can be securely lowered from.
t Lower the cover plate far enough that the stirrer elements and the sparger pipe
can be easily maintained and optionally converted, if necessary.
Or lower the cover plate to the floor or onto a table suitable for transportation
(carriage).
t Gently place the cover with the stirrer shaft to the side so as not to damage the
shaft, still-mounted stirrers or the sparger pipe.
t You can now remove the lifting system sleeve and take away the cover.
7.3.2 Lowering the Cover t Swivel lifting system into position [1] and secure with locking pin.
Plate onto the Vessel t Place table with cover plate under the lifting system (again) so that the cover plate
can be lifted upward.
t Lower the lifting system so that the sleeve can be fitted.
t Screw the lifting system sleeve into the stirrer shaft adapter. Gently turn the crank
to lift the cover. The second person should hold and guide the cover so that no
part (cover, stirrer shaft, stirrer elements or sparger pipe) can run into the outside
of the vessel or fittings / valve assemblies installed on it.

If the bottom edge of the sparger pipe / stirrer shaft lie above the vessel flange,
the lifting arm can be pivoted:
t Take the locking pin out of position [1] and lock it into position [2] after tilting.
t Carefully turn the crank to lower the cover plate. The second person should guide
the cover so that the attachment parts (stirrer shaft, stirrer and sparger pipe) do
not strike the inside of the vessel.
t Lower the cover plate onto the vessel flange and align it so that the cover's screw
connections are above their threaded holes in the flange. When lowering and
aligning the cover plate, ensure that the cover plate's O-ring sits properly in its
groove and does not bend or stick out.
t Screw in cover connections. They should be easy to screw in (without jamming).
If not, slightly lift the cover plate once again and correct the alignment of the
vessel flange.
t Loosen the lifting system sleeve from the stirrer shaft coupling and turn it up
again. If necessary, swing the lift arm into its resting position and secure (locking
pin [1] in position [Ä]).
t Install the required fittings in the side probe port N1 (N2, where applicable) or
mount on the cover plate and establish all the necessary connections (supply,
exhaust, motor, intakes and gas lines, condensate / cooling water).
7.4 Disassembly / Assembly For example, the cover must be removed to perform the following:
of the Cover Plate − Cleaning the vessel
− Conversion of assembly parts in the vessel (aeration, stirrer)
Depending on the design of the vessel, the cover is connected to the vessel with
hexagonal screws or locking bolts.
Prior work
Danger of injury by escaping materials!

When the cover is opened, gaseous and liquid materials may escape under high
pressure and e.g. cause injury to eyes.
Ensure that the culture vessel is depressurized.
Turn the unit off and secure it against reactivation before removing the cover.
Danger of crushing by vessel cover!

Extremities may be crushed during assembly and disassembly.
Only hold the cover using the handles provided.
Two people working together should lift the cover.
Dismounting/mounting with lifting accessories:
Use only the lifting accessories provided to lift the cover.
Attach the lifting accessories only to the fastening points provided.
t Disassemble the motor [¨ see Section “7.2 Assembly / Disassembly of the Motor”].
t Loosen the connections of parts connected to the periphery of the frame, such as
the gas supply connection on the supply air filter and the exhaust air assembly
with the exhaust air filter.
t Remove the parts installed in the cover plate if these interfere with disassembly or
can be damaged.

Important information
Loose parts such as clamps and gaskets must be stored securely.
t Remove all components that project deep into the vessel (e.g. sensors that are
screwed in, etc.).
t Turn the device off at the main switch.
Cover Lifting System

If the device is equipped with a cover lifting system, follow the instructions in
[¨ Section “7.3 Cover Lifting System”].
Disassembly of the Cover Plate

t Loosen the cover screws and carefully lift the cover plate with 2 people.
t Place the cover carefully flat on the floor.
y The vessel is now accessible for cleaning and maintenance work.
Assembly of the Cover Plate

t Check the O-ring on the cover plate. It must be free of adherent dirt and
undamaged. Clean or replace it.
We recommend slightly greasing the O-ring with silicone grease.
t Replace the cover plate on the vessel flange and align it carefully.
The holes in the cover and vessel flange should be exactly superimposed and the
screws should snap in easily. Firmly tighten the locking bolts in a diagonal pattern.
t In reverse order of disassembly, reconnect the components installed in the cover
and peripheral connections.
7.5 Installation of the Stirrer t Lift the cover plate with the stirrer shaft off of the vessel [¨ see Section “7.4
Disassembly / Assembly of the Cover Plate”].
Assembly Information
t Establish the installation height of the stirrer elements on the stirrer shaft.
The bottom-most stirrer should be at the lower end of the stirrer shaft.
The top-most stirrer should still be immersed in the medium even with the smallest
working volume.
The middle stirrer should be centered between them.
Depending on the accessories installed in the vessel, the stirrer should be arranged so
that optimum medium mixing results can be achieved at all speeds and no liquid
streams can form.
Installation of the stirrer

1. Check the position and attachment of the installed stirrer. Correct the position if
necessary, and affix the stirrer with the mounting screws.
2. If you want to remove or replace the stirrer elements, loosen the mounting screws
and remove the stirrer from the shaft. Slide the new stirrer to the desired height
and tighten the mounting screws.

7.6 Cleaning
Inappropriate cleaning agents pose the danger of corrosion and damage to the
device and the culture vessel.
− Avoid strongly caustic or alkaline and/or chloride-containing detergents.
− Avoid solvent-based cleaning agents.
− Ensure that the cleaning agents used are compliant materials.
Observe the safety instructions for the cleaning agents. The use and disposal of
cleaning agents, and water containing the same, may be subject to legal or
environmental protection regulations.
7.6.1 Cleaning the Device

Danger of electric shock due to foreign liquids and objects!
Device must not be cleaned with high-pressure cleaners etc. or direct water stream.
t Clean the device housing with a slightly damp cleaning cloth; for more severe
contamination, use a mild soap.
t Clean the operator display with a slightly damp, lint-free cleaning cloth; for more
severe contamination, use a mild soap.
Be sure not to scratch the device or operating display. Future contamination is

otherwise more difficult to remove.
7.6.2 Cleaning the Control

Danger of corrosion and damage to the vessel and attachments.
Tower, Vessel, and
Avoid strongly caustic or alkaline and/or chloride-containing detergents.
Attachments
It may be necessary to lift off the culture vessel cover and mechanically clean the
culture vessel and attachments.
Disconnect the system from power before removing the cover [¨ see Section “7.4
Cleaning and maintenance intervals depend mainly on how much the vessel and
equipment are used and contaminated (by aggressive constituents of the medium,
such as acids and alkalis, by substances formed during cell growth, by contamination
with media components, etc.).
1. Check whether it is sufficient for the process to rinse the vessel, attachments, and
accessories with water.
− If contaminated by organic substances, you can clean glass parts with
commercially-available special laboratory glass cleaners (e.g. RBS from ROTH,
NEODISHER, or similar) in a warm solution.
− You can mechanically remove stubborn stains, inorganic deposits can be
removed with dilute sodium hydroxide solution (or similar).

2. You can clean metallic parts (cover plate, etc.) mechanically, if needed with the
help of mild detergents or alcohol. Make sure that you do not cause any
scratching.
− If you use cleaning agents that may affect the subsequent process, the vessel
and attachments must be rinsed subsequently.
3. You can carefully clean seals and O-rings mechanically. Chemical cleaning agents
can attack the material and adversely affect the sealing effect. The seals and
O-rings must be replaced if adherent dirt cannot be removed after the application
of silicone grease.
7.6.3 Intermediate Cleaning 1. As required, dismount attachments from the cover to gain access to the interior of
After Processes the vessel.
If necessary, dismount the cover plate.
2. Rinse the vessel carefully with water.
3. Check the vessel attachments. If contamination is still present, dismount and clean
the electrodes and other attachments. Then install them again.
4. Fill with demineralized water until at least the pH and DO electrodes are covered.
The pH and DO electrodes may not dry out; otherwise they will require lengthy service
before reactivation.
7.6.4 Basic Cleaning and When putting the unit out of operation for longer intervals, you should disassemble
Storage all equipment and fittings mounted on and in the vessel.
1. Empty the vessel completely. Disassemble the cover plate [¨ see Section “7.4
2. Remove all electrodes and accessories and clean all parts.
3. Especially check all gaskets and O-rings. Replace them if damaged (if necessary,
even if pressure marks or hairline cracks can be detected) or if tough contaminants
cannot be removed. Grease the seals with a silicone lubricant.
4. Store the parts as recommended for the individual case. For storing the electrodes,
observe the information in any related documents or the manufacturer’s
information.
7.7 Maintenance Instructions

and Function Tests
7.7.1 Measures to Take t After maintenance tasks, carry out a visual inspection of the unit and ensure that
after Maintenance all connectors and connection points are tight.
t Tighten the screws of the tri-clamp finger-tight.
t Check the pneumatic connections of the valve.
t Carry out a pressure test of the unit if necessary [¨ see Section “6.13.1 Sterile Test
and Pressure-hold Test”].

7.7.2 Device Maintenance Maintenance performed by the user is restricted to the following tasks:
− Work on the vessel is limited to the installation and removal of the cover plate for
access to the interior, cleaning the vessel, the installation, removal or adjustment
of the components installed in the vessel and the installation and removal of
electrodes and accessories. Maintaining the pH, DO or Redox sensors as per the
manufacturer's | supplier's specifications.
− Checking and replacing parts subject to wear as well as disposables, e.g., glass
vessels, filters, tubing and seals with identical components with the same
specifications [Spare parts list].
− Replacing O-rings, seals, filters, hoses and disposable items, e.g. inoculum
membranes.
The internal modules of the device, and the safety equipment, pump modules,
drive motors and stirrer shaft couplings, should only be serviced by qualified and
correspondingly authorized service personnel.
Any servicing instructions for internal equipment, electrical modules and safety
equipment contained in this manual and the technical documentation must be
forwarded to the servicing personnel.
7.7.3 Maintenance Intervals The cyclical maintenance of the equipment and its components depends on the
process conditions and frequency of use and duration. Have the components checked
by a specialist during inspections according to legal and|or internal company
regulations, but at minimum according to the following intervals. The following
tabular overview is not binding and must be adapted to individual conditions
(shorter intervals).
Sartorius accepts no liability for incorrectly scheduled maintenance intervals.

Component Activity Before every After 10-20 If unsterile 1 + yearly
process autoclave cycles
Culture vessel
Leak test Pressure-hold test
+ +
Tightness test
Connections between device
and culture vessel, air, water
Leak test Visual inspection +
Temperature control system
Leak test Visual inspection +
Tapping septa
Replace +
O-rings
Visual inspection,
+
replace if needed
Replace + + +
Intake and exhaust air filters
Filter cartridges Integrity testing +
Replace + +
Replace + + +
Holding bottles,
sampling bottles
Visual inspection,
+
replace if needed
Gaskets / ventilation filters Replace + +
Rotating mechanical seal
Check for contamination and Visual inspection
+
damage
Peristaltic pumps
Pump hoses Visual inspection,
x
replace if needed
Safety valves
Culture vessel Functional test after trigger-
+
ing, replace if needed
Temperature control system Functional test after trigger-
+
ing, replace if needed

Component Activity Before every After 10-20 If unsterile 1 + yearly
process autoclave cycles
Sensors Visual inspection,
replace if needed
pH sensor Calibration, visual inspection
+
for damage
DO sensor Calibration, visual inspection
+
for damage
Membrane body / electrolyte Visual inspection,
+
(Clark sensors) replace if needed
Sensor cap Visual inspection,
+
(optical O2 sensor) replace if needed
Foam sensor Calibration, visual inspection
+
for damage
Level sensor Calibration, visual inspection
+
for damage
Temperature sensor Calibration, visual inspection
+
for damage
Plugs / contacts / lines
Visual inspection +
Maintenance according to
maintenance schedule
Maintenance and functional test Only to be carried out by
according to maintenance report Sartorius experts
+
Contact Sartorius Stedim
Service.

7.7.4 Seals
7.7.4.1 O-ring Gaskets O-ring gaskets (2) seal installed attachments such as sensors and intake systems to
the vessel.
O-ring gaskets are consumable materials and must be regularly checked for damage
and wear and replaced if necessary.
A list of all consumable materials is found in the consumable material list in the
“Overall documentation” folder.
2
Changing O-ring gaskets
Remove the corresponding attachment from the vessel and carry out a visual
inspection of the O-ring gasket.
t Replace a damaged or no longer taut gasket.
t If necessary, moisten the new O-ring gasket with lubricant.
The lubricant must be approved for operation with oxygen.
t Place the attachment into the corresponding port and tighten it finger-tight.
7.7.4.2 Tri-clamp Gaskets Tri-clamps (1) seal the connections between pipelines and the different functional
modules and ensure reliable leak tightness.
To ensure the functionality of the tri-clamps, the gaskets must be checked regularly
for damage and wear. Damaged or worn gaskets must be replaced.
1
Tri-clamps are installed in different sizes. A list of all consumable materials is found in
2 the consumable material list in the “Overall documentation” folder.
Changing tri-clamp gaskets

t Loosen the wingnut (2) of the tri-clamp and remove the tri-clamp from the
3 connection.
t Replace the gasket (3).
t When reassembling, ensure that the gasket is in the groove for the connection
nozzle.
The diameter of the gasket opening (d2) must be greater than the cross-section of the
line (d1), since if the tri-clamp gasket is tightened too firmly the sealing material will
be pressed into the line.
Flange Gasket

7.7.5 Replacing Intake and The intake and exhaust air filters must be replaced before each process, after faults,
Exhaust Air Filters and during periodic maintenance [¨ see Section “7.7.3 Maintenance Intervals”].
Changing filters (intake air path)

t Release the quick release coupling (1).
1
t Unscrew the connection (2) and the top part of the filter housing (3).
t Remove the filter.
t Check the O-seal for damage and replace it if necessary.
t Insert the new filter (4) into the filter housing.

t Put on the top of the filter housing.
t Carefully tighten the screw connection (2) finger-tight.
Changing filters (exhaust air path)

t Loosen the screw (1) on the tri-clamp on the filter housing.
t Remove the tri-clamp (2) and the top part of the filter housing (3).
t Remove the filter.
t Check the tri-clamp gasket for damage and replace it if necessary [¨ Section
3 “7.7.4 Seals”].

t Insert the new filter (4) into the filter housing.
t Place the gasket onto the edge of the lower filter housing and place the top filter
housing onto it.
4
t Check for correct seating of the gasket (5) between the top and bottom filter
housing.
t Place the tri-clamp around the housing flange and tighten the clamping screw
finger-tight.
5
7.7.6 Replacing the View The view glass lamp (1) is located on the view glass in the cover port.
Glass Lamp
t Remove the cylinder with the view glass lamp from the port.
t Screw the cylindrical halves (2) of the view glass housing apart.
t Remove the lamp from the cylinder half.
t Replace the bulb.
2 t Reassemble the view glass housing in reverse order.
t Place the view glass lamp onto the port of the view glass.
Fig. 7-1: Replacing the bulb of the view glass lamp

7.7.7 Sensors Depending on configuration, sensors may be installed in the cover and the lower part
of the vessel [¨ Chapter “3. Device Overview”]. The ports for sensors are available in
d 19 mm and d 25 mm variants.
12 mm sensors with PG13.5 thread, such as sensors for pH, DO and Redox, must be
screwed into an attachment nozzle in the culture vessel before installation.
t Insert the sensor carefully into the attachment nozzle until it hits the stop.
t Turn the sensor in the clockwise direction until it is screwed in to finger tightness.
t Install the sensor and the attachment nozzle into a d 25 mm side port.
Fig. 7-2: Attachment nozzle 25 mm on PG 13.5
Danger of burns!
Make sure that the sensors are properly installed in the installation port.
Incorrectly installed sensors can be pushed out of the port during the sterilization
process.
Calibration
The following sensors are calibrated by the operator of the unit:
− pH sensor
− DO sensor
− Turbidity sensor
− Redox sensor
The following sensors are calibrated by Sartorius Stedim Service:

− Pressure sensor
− Temperature sensor
O-ring gaskets for sensors

t Before calibration and the functional test, check the O-ring gaskets and replace
them if necessary [¨ see Section “7.7.4.1 O-ring Gaskets”].

7.7.7.1 pH Sensor pH sensors are subject to age and wear due to:
− e.g. thermal influences during sterilization
− Chemical reactions with the culture medium
− Or deposits such as protein on the diaphragm.
Symptoms include poorer triggering or reduced sensor slope. If the diaphragm

is subject to organism growth, the pH measurement value will differ between a
standing culture medium and agitated culture medium.
The functional test of the pH sensor is restricted to checking the zero point and slope
after calibration. Follow the instructions in the manufacturer documentation for the
component.
t Make sure that the sensor cable is connected.
Calibrating the pH sensor

The pH sensor must be calibrated while not attached to the vessel. You can find
instructions for the calibration and parameters for pH measurement in [¨ Part B:
Digital Measurement and Control System].
Installing in the vessel

t After calibration, insert the pH sensor into a d 25 mm side port (1).
t Carefully tighten the screw connection finger-tight (2).
t Connect the electrode to the socket “pH” of the supply unit.
If the cap nut (3) is not correctly tightened, the sensor will not be sealed and the
vessel cannot be filled or pressurized.
Storing the pH sensor

pH sensors should always be stored with an irrigation cap filled with 3M KCl solution
or storage solution.

7.7.7.2 DO Sensor (pO2) t Check the function of the DO sensor before each new process.
t Make sure that the sensor cable is connected.
Calibrating the DO sensor

You can find instructions for the calibration of the DO sensor and its parameters in
[¨ Part B: Digital Measurement and Control System].
Calibration includes the setting of the electrode zero point and the measurement of
slope.
If using Clark sensors:

Before calibration, the sensor must be polarized for approx. 6 hours. You must repeat
the polarization if the sensor is disconnected from the amplifier for about 10 minutes
[¨ see specifications for the sensor in the manufacturer's documentation].
Optical DO sensors must not be polarized.
Calibrating the zero point:

t Aerate the culture medium using nitrogen until the dissolved oxygen is fully
eliminated.
Calibrating slope:
t Aerate the medium with air or gas mixture.

t After calibration, insert the DO sensor into a 25 mm side port d 25 mm (1).
t Carefully tighten the screw connection finger-tight (2).
If the cap nut (3) is not correctly tightened, the sensor will not be sealed and the
vessel cannot be filled or pressurized.

7.7.7.3 Antifoam and The level sensor and the antifoam sensor are identical in construction. The instruc-
Level Sensor tions for the installation, maintenance, and handling are the same for both sensors.

The sensor is installed in an adapter (2) in the cover port.
The stay bolt (3) on the tip of the sensor prevents overpressure in the vessel from
forcing the sensor out of the adapter if the cap nut (6) is not correctly tightened.
The sensor may be overgrown by cells, cell residue, or medium residue. This coating
may hinder the conductivity measurement. If necessary, clean the sensor. Replace
damaged O-rings.
t Check the sensor and O-rings (4) on the adapter.

t If the adapter (2) is not yet installed, twist the stay bolt (3) out of the sensor shaft
(1). Push the adapter over the shaft. Twist the stay bolt back in.
t Insert the sensor and adapter (2) into the d 19 mm cover port and tighten the
adapter.
t Push the sensor to the desired height and fasten with the cap nut (6). The clamping
cone (5) fastens the sensor at the adjusted height.
t Connect the cable of the sensor to the corresponding connectors on the control
unit.
The sensor may not lie too closely above the medium. This avoids contact with the
medium at high agitator speeds or intensive aeration.
You can see the installed position of the sensor in the [¨ Chapter “3. Device
Overview”].
7.7.7.4 Turbidity Sensor Calibrating the turbidity sensor

Calibrate the turbidity sensor when it is not installed in the vessel. You can find
instructions for the calibration and parameters in [¨ Part B: Digital Measurement
and Control System].

t After calibration, insert the turbidity sensor into a d 25 mm side port. To avoid
dead spaces and permit reliable sterilization, the O-ring must be close to the sensor
in the medium.
t Carefully tighten the screw connection finger-tight.
t Connect the sensor to the corresponding connector on the control unit (see the
technical documentation in the overall documentation).
If the cap nut is not correctly tightened, the sensor will not be sealed and the vessel
cannot be filled or pressurized.

7.7.7.5 Redox Sensor The Redox calibration includes a functional test of the Redox sensor. The functional
Functional Test test of the Redox sensor is carried out prior to installation in the culture vessel, i.e.,
before sterilization.
Calibrate the Redox sensor when it is not installed in the vessel. You can find
instructions for the calibration and parameters in [¨ Part B: Digital Measurement
and Control System].

t After calibration, insert the Redox sensor into a slanted d 25 mm side port.
To avoid dead spaces and permit reliable sterilization, the O-ring must be close
to the port or sensor adapter in the medium.
t Carefully tighten the screw connection finger-tight.
t Connect the sensor to the corresponding connector on the control unit (see the
technical documentation in the folder “overall documentation”).
If the cap nut is not correctly tightened, the sensor will not be sealed and the vessel
cannot be filled or pressurized.
After installing all other sensors and attachments into the side ports, fill water into
the vessel until the sensors are covered.
This will prevent the sensors from drying out.
7.7.8 SACOVA Valve If using a SACOVA valve to transfer liquids, it is practical to clean the valve after each
work process by careful rinsing. The valve may need to be disassembled in the case of
encrustations due to adhering media residues.
The SACOVA valve is only functional if it closes absolutely tightly in its closed
position. It should therefore be checked for leaks regularly.
If, due to unavoidable aggressive media or media concentrations, the O-rings

(standard: EPDM 3957 and FDA approval) or the seating surfaces of the O-rings in
stainless steel (material no.: 1.4435 = 316 L) are attacked, the tightness should be
checked after each operation.
This can be easily seen by submersion in a water bath with compressed air!
Disassembly and assembly of the SACOVA valve

1. Remove silicone hose
2. Remove the locking ring
3. Remove the knurled nut
4. Remove clamp
5. Remove with locking ring
6. Carefully remove the valve tappet from the valve sleeve.
7. Check all O-rings and, if damaged, replace them. Lightly grease O-rings with
suitable O-ring grease to prevent them from sticking to contact areas.
8. Following the disassembly steps in reverse order to assemble the SACOVA valve.

7.7.9 Inoculation Fittings If inoculation kits have already been used, be sure to test the inner tube and
and Septa inoculation needle (3).
t Remove any adhering residue of media previously transferred.

t Test the O-ring and replace it if necessary.
From time to time, check the fiberglass filling (7) in the sterile sleeve.
t To do so, unscrew the screw cap (6) and the hose nozzle.
t Replace the fiberglass if it is wet or dirty.
Pos. Description
1 Inoculation fitting
2 Cap nut
3 Inoculation needle
4 Hose nozzle
5 Sterile sleeve
6 Screw cap
7 Glass fiber filter packing
Fig. 7-3: Inoculation fitting
t Replace used (perforated) inoculation membranes before each new process.
t Check all O-rings and replace it if it is porous, has pressure marks, or is damaged.
Fig. 7-4: Septa

7.7.10 Internal Spinfilter Clean spin filter
The spin filter can be cleaned as follows:
− Use an appropriate laboratory dishwasher with sufficient rinsing chamber size.
− If the filter is contaminated with cell residue or components of the culture
medium, clean it with a brush if necessary.
− Clean the spin filter overnight in a solution of 0.5-1 N NaOH at 80 ° C.
Then neutralize the NaOH solution and rinse the spin filter carefully with deionized
water.
− Clean the spin filter for 30 minutes at 50 ° C in an ultrasonic bath.
When using other cleaning agents, ensure that these do not affect the culture process
and thoroughly rinse the spin filter with deionized water afterwards.
Harvesting pipe spin filter
3
4
Pos. Description
1 Dummy Plugs
2 Septum holder
3 Inoculation nozzle
4 Septum
5 O-ring
6 O-ring
7 Harvesting pipe

Assembly position of the spin filter and the harvesting pipe spinfilter
The spin filter must be mounted on the stirrer shaft so that the top edge is above the
culture medium. The culture medium may not flow over the edge into the inner space
of the spin filter.
Where necessary, determine the respective maximum and minimum filling levels at
settings for maximum stirrer speed and maximum aeration.
Note that the filling volume can vary due to aeration, stirring and, where applicable,
additional equipment.
Cell-free medium is harvested using the harvesting pipe spin filter. The harvesting
pipe spin filter must be installed in a cover port (19 mm in diameter) and positioned
in the inner portion of the spin filter [¨ see Section “3.5 Culture Vessels”]. The lower
end of the harvesting pipe should reach to the bottom of the spin filter.
Mounting the spin filter harvesting pipe spin filter

t Remove the stirrer driver from the stirring shaft [¨ see Section “7.2
Assembly / Disassembly of the Motor”].
t Remove the cover plate and set it aside so that you can work on all areas.
t Guide the harvesting pipe SF with the adapter into the appropriate septum of the
cover port and screw it in place.
t Position the harvesting pipe so that it is in the interior of the mounted spin filter.
t Slide the spin filter onto the stirrer shaft.
Position the spin filter so that the lower end of the harvesting pipe SF is in the
lower region of the spin filter.
t Attach the spin filter with the screw pin to the stirrer shaft.
t Mount the disk impeller on the stirring shaft below the spin filter.
t Insert an inoculation membrane into the holder.
t Screw in a dummy plug.
t Position the cover plate so that the harvesting pipe SF projects into the interior of
the spin filter.
t Install the remaining culture vessel components and attach the cover plate to the
culture vessel.

7.7.11 STT Quick Connector − The slit membrane inside the STT coupling part can be punctured repeatedly, but
should only be used for one particular process. The membrane slit can be soiled or
damaged when punctured multiple times, making it unable to sterile self-seal.
− The O-ring on the plug and|or on the dummy plug of the coupling part wears out
slowly and must only be replaced in case of visible roughness or damage.
Spare and Wearing Parts
Order no. Component, specifications

8809267 Slit membrane for sealing couping, item no. 8809240 / 8809410
− 20-piece package
39971414 Silicone hose 3.2 + 1.6 mm (interior diameter + wall thickness)
− Minimum order 10 m
39120945 O-ring 7.65 + 1.78, made from EPDM, FDA approved
− 10-piece package
7.8 Assembling and Adjusting The sparger pipe is pre-assembled and need not be removed for inspection and
the Sparger Pipe cleaning, e.g. if the holes in the injection ring are clogged by media residue or growth
from a past process.
Disassembly is only required if another sparger pipe, e.g. with micro-sparger, is to be
installed.
The sparger is attached to the adapter for in-situ sterilization of the supply filter.
After removing the cover plate, lift the sparger pipe out of the culture vessel.
Loosen the screw to remove it from the cover plate.
7.9 Securing the Weigh Cells If the bioreactor is equipped with a weighing system, a total of 3 weigh cells (on top
while Moving the Unit of which the vessel rests) are mounted to the left, right and rear of the base frame (4),
[¨ see Section1 “Fig. 7-5: Removing the transport lock”].
− The lugs (1) on the brackets (2) are used for safe transport and must be removed
before starting up operations. During weighing mode, the force is directed over
the stay bolts (3) and the holders (6) onto the weigh cells.
− The vessel is connected to the base unit via flexible hose connections.
− The weighing system is operated using the associated measurement and control
system operating menu [¨ see Part B: Digital Measurement and Control System].
Shipping Instructions
During transport, the vessel is mounted with the lugs (1) in the brackets (2) to the left
and right of the base frame.
The stay bolts (3) left, right and rear are lifted out of the weigh cell holders (6) at
“d = approx. 3 mm”. This is necessary to avoid damage during transport of the weigh
cells.
1. Carefully transport the bioreactor to the installation location with the vessel
installed. Use suitable transport equipment.
2. Carefully horizontally align the bioreactor after installation at the workplace.
You can compensate for unevenness in the floor with the adjustable legs.
1 Construction and arrangement of transport locks and suspension may vary from the version shown here
for different bioreactors and culture vessels.

Remove transport lock
1. Loosen the screws on the lugs (1) on the right and left of the vessel.
2. Lift the boiler slightly so that you can remove the lugs (1).
3. After removing the lugs, lower the vessel slowly until the stay bolts (3) are resting
in the holders (6).
Sudden lowering of the vessel can push out the stay bolts and damage the weigh
cells!
4. Please retain the transportation locks. Install them again before transporting the
bioreactor to a different location.
5.
Front view, transport condition Side view, transport condition
Front view, operating state Side view, operating state
Fig. 7-5: Removing the transport lock
Pos. Description Pos. Description

1 Lug 4 Weigh cells
2 Mounting bracket for the vessel 5 Mounting bracket for weigh cells
3 Stay bolts 6 Holder for stay bolts

8. Disruptions
8. Disruptions
8.1 Safety Instructions
Contact with parts under voltage can be life-threatening.
− Work on the electrical equipment of the device may only be carried out by a
competent electrician.
− Before any work, turn the device off and disconnect it from power.
− Always make sure any electrical equipment is disconnected from AC power before
performing any maintenance, cleaning or repair work.
Danger of limbs being pulled into the rotation pump and crushed!
− Allow only qualified and authorized personnel to work on the device.
− Disconnect the device from power when performing maintenance and cleaning
tasks.
Danger of burns upon contact with hot surfaces!

− Avoid contact with hot surfaces like vessels, motor housings, and pipelines carrying
steam.
− Let the vessels/culture vessels cool before troubleshooting.
8.2 Troubleshooting Always proceed according to the following steps when faults occur on the device.
1. Switch off the device and unplug it from the power supply (pull power plug) if the
fault (e.g. smoke or odors, abnormally high surface temperatures) represents a
direct danger to personnel or property.
2. Inform management on site about the fault.
3. Determine the cause of the fault and remedy it before switching the device back
on [¨ see Section “6.2 Switching the Device On | Off”].
If the fault cannot be remedied, please consult your Service Center [¨ Section “1.2
Service Center”].
8.2.1 Process-related
Faults in the operating sequence are displayed as alarms on the operator terminal.
Faults
To correct these process-related faults, read the [¨ Part B: Digital Measurement and
Control System, Chapter “20. Appendix”].
8.2.2 Hardware-related
Danger of injury if personnel qualifications are insufficient!
Faults
Improper use can lead to significant personal injury and| or property damage.
It is therefore important that all troubleshooting activities be carried out by qualified
personnel.
110 Disruptions
8.2.3 Fault Table Contamination Possible causes Corrective measures
“Contamination”
Generalized and Insufficiently autoclaved Check the autoclave settings.
widespread, even culture vessel.
Increase the autoclaving time.
without having
inoculated the culture Perform sterility tests using test
(during the sterility spores.
test phase)
Air inlet line or air inlet Replace the tubing.
filter defective.
Check the filter and replace
if necessary.
Generalized and Seals on the culture vessel Carefully check the integrated
gradual (even without or the integrated parts.
inoculating the components are damaged
If suspected to be damaged
culture) (e.g. hairline cracks).
(rough, porous surfaces or
dents), replace seals.
After inoculation, Contaminated inoculation Take control samples of the
wide-spread culture. inoculation culture and test
inoculated culture medium
Non-sterile inoculation
from the vessels (e.g. on test
equipment
nutrient solutions).
Incorrect inoculation Check the inoculation
procedure.
Carefully practice the
inoculation process.
Air inlet filter or Check the filter(s) and replace,
connection not sterile or if necessary.
defective
Replace the connection line.
During the process, Air inlet filter or Check the filter(s) and replace,
rapid connection not sterile or if necessary.
defective
Accidental or unauthorized Take organizational measures
manipulation of at your work area to prevent
components tampering with the equipment.
During the process, Seals on the culture vessel If possible, continue process
gradually or the integrated up to the end point. At the end
components are damaged of the process, dismantle the
(e.g. hairline cracks or vessel and carefully check the
porous) integrated components.
If suspected to be damaged
(rough, porous surfaces or
dents), replace seals.
Exhaust air filter(s) or Check the filter(s) by performing
connection has become a sterility test, if possible, and
unsterile or is defective replace if necessary.
(contaminated from the
exhaust air line).
Disruptions 111
We recommend that you perform a sterility test before each process.
Duration 24 - 48 h.
Conditions for a Sterility Test:

− The culture vessels must be filled with the scheduled culture medium or a suitable
start medium and autoclaved in accordance with the specifications.
− All of the scheduled components, peripheral devices, correction media supply
systems and sampling systems must be connected to the culture vessels.
− The system must be set to the scheduled operating conditions (e.g. temperature,
stirrer speed, aeration).
8.2.4 Troubleshooting Table The counter cooling system does not work or does not provide sufficient cooling
“Counter Cooling power.
System”
Problem Possible causes Corrective measures
The cooling water The laboratory’s supply line If all other potential causes
is not being fed into is blocked or the valves of can be excluded (see below),
the system the cooling water feed are contact customer service.
defective.
The cooling water supply Check the water hardness
valve does not work or the (no more than 12 dH).
non-return valve has
Check the non-return valve.
become stuck because of
contaminated cooling Feed clean cooling water into
water or scale deposits. the system (if necessary, install
a pre-filter).
Insufficient cooling Flow rate too low The minimum operating
power temperature is around 8° C
Cooling water tempera-
above the cooling water
ture too high
temperature.
If necessary, install an upstream
cooling device.
8.2.5 Troubleshooting Table Aeration or ventilation does not work or is insufficient.

“Aeration and
Ventilation” Problem Possible causes Corrective measures
Air inlet line blocked Air inlet filter blocked. Check the air feed (dry and free
of oil and dust).
Install a pre-filter, if necessary.
The gas or air Hose kinked or Check the hose and filter and,
supply is blocked or disconnected. if necessary, fit new sterile
decreases suddenly Exhaust air filter blocked filters.
(e.g. as a result of moist air
and the formation of
condensate, or foam).
112 Disruptions
9. Disposal
9. Disposal
9.1 General Notes The packaging is to be taken to a local waste disposal site if no longer required.
The packaging is made of environmentally friendly materials that can be used as
secondary raw materials.
The equipment, including accessories and batteries, should not be disposed of as

regular household waste.
EU legislation requires its Member States to collect electrical and electronic
equipment and dispose of it separately from other unsorted municipal waste so that
it may be recycled.
In Germany and several other countries, Sartorius Stedim Biotech GmbH itself
assumes responsibility for the return and legally compliant disposal of its electronic
and electrical products. These products may not be placed with household waste or
brought to collection centers run by local public disposal operations – not even by
small commercial operators. For disposal in Germany and in the other member
nations of the European Economic Area (EEA), please contact our local service
technicians or our Service Center in Goettingen, Germany:

August-Spindler-Strasse 11
Telephone +49.551.308.0
Fax +49.551.308.3289
In countries that are not members of the European Economic Area (EEA) or where no
Sartorius subsidiaries or dealerships are located, please contact your local authorities
or a commercial disposal operator.
Prior to disposal and/or scrapping of the equipment, any batteries should be removed
and disposed of at local collection points.
Sartorius will not take back equipment contaminated with hazardous materials
(ABC contamination) – either for repair or disposal. Detailed information with service
addresses for returning your device for repair or disposal can be found on our website
(www.sartorius.com) or requested from a Sartorius Service Center.
9.2 Hazardous Materials The device does not contain any hazardous materials that require special disposal
measures.
The cultures and media (e.g. acids, bases) used during fermentation or cultivation
process are potentially hazardous materials that might cause biological or chemical
hazards.
Note in accordance with European directive on hazardous substances!
According to the EU directives, the owners of devices that come into contact with
hazardous substances are responsible for properly disposing of these devices and to
declare such devices when transporting them.
Corrosion
When using corrosive gases, the fittings must be chosen accordingly, (e.g. made of
stainless steel instead of brass). To change such fittings, please contact Sartorius
Stedim Service.
We do not accept any responsibility for operating faults and defects resulting from
the use of unsuitable gases.
Disposal 113
9.3 Decontamination Sartorius Stedim Systems GmbH has a duty to protect its staff from hazardous
Declaration substances. When returning devices and device components, the sender must enclose
a decontamination declaration as proof of compliance with the safety regulations
governing the area of application for which the devices were used.
This declaration must detail the microorganisms, cells and media that the device has
come into contact with and the measures taken to disinfect and decontaminate it:
− The recipient (e.g. Sartorius Stedim Service) must be able to read this
decontamination declaration before opening the packaging.
− This document contains an attachment with a decontamination declaration form
in [¨ Section “10.5 Decontamination Declaration”].
To create additional copies, simply copy the attached form or request additional
forms from Sartorius Stedim Systems GmbH.
Danger of severe injury from improperly performed work!

The disassembly and disposal of the device may only be carried out by qualified
personnel.
Warning of dangerous electrical voltage!

Work on the electrical equipment may only be carried out by a competent electrician.
9.4 Decommissioning Carry out the following work steps for the disassembly of the device:
the Device t Empty the culture vessel, pipelines, and hoses of all culture media and additives.
− Carry out a cleaning of the entire device.
t Carry out sterilization of the entire device.
t Turn the device off via the device's main switch and secure the device against
being turned back on.
t Disconnect the device from power and the supply lines.
9.5 Disposing of
Danger of severe injury due to ejected or falling parts!
the Device
When disassembling the device, pay particular attention to those components that
contain parts under mechanical tension that could spring out during scrapping,
leading to injury. There is also danger due to moving parts and falling objects.
− The device may only be disassembled by qualified personnel.
− Disassemble the device carefully and in a safety-conscious manner.
− Wear the following personal protective equipment during work
[¨ see also Section “2.15 Personal Protective Equipment”]:
− Safety gloves
− Protective work clothes
− Safety shoes
− Safety glasses.
t Disassemble the device until all device parts have been assigned to a material
group and can be appropriately disposed of.
t Dispose of the device in an environmentally friendly manner. Follow the
regulations applicable in the local region.
114 Disposal
10. Appendix
10. Appendix
10.1 Specifications
Culture vessel volume 5 L 10 L 15 L 20 L 30 L

Dimensions [W + H + D] [“] 35.4 + 51.2 + 27.6 39.4 + 74.8 + 29.5 39.4 + 74.8 + 29.5 39.4 + 74.8 + 29.5 39.4 + 74.8 + 29.5
[m] 0.9 + 1.3 + 0.7 1.0 + 1.9 + 0.75 1.0 + 1.9 + 0.75 1.0 + 1.9 + 0.75 1.0 + 1.9 + 0.75
Required door dimensions for [“] 31.5 + 51.2 31.5 + 78.7 31.5 + 78.7 31.5 + 78.7 31.5 + 78.7
installation [W + H] [m] 0.8 + 1.3 0.8 + 2 0.8 + 2 0.8 + 2 0.8 + 2
Weight (approx.) [kg] 130 210 215 215 230
Environmental temperature | Relative humidity <80 % for temperatures up to 31 °C (87.8°F), decreasing linearly < 50 % at 40 °C (104°F).
(non-condensing)
Supply lines Specifications Max. Culture vessel volume

flow rate 5 L 10 L 15 L 20 L 30 L
Process | control air MO | 4 – 6 barg | 58 – 87 psig, preset, [L/min] 7.5 | 15 | 23 | 30 | 45 |
CC sparger | overlay Class 2 (ISO 8573-1) 0.5/5 1/10 1.5/15 2/20 3/30
O2 MO sparger | 4 barg | 58 psig, preset, free from particles [L/min] 7.5 | 15 | 15 | 30 | 45 |
CC sparger 0.5 1 1.5 2 3
CO2 MO sparger | 4 barg | 58 psig, preset, free from particles [L/min] N|A N|A N|A N|A N|A
CC sparger 0.5 1 1.5 2 3
N2 MO sparger | 4 barg | 58 psig, preset, free from particles [L/min] N|A N|A N|A N|A N|A
CC sparger 0.5 1 1.5 2 3
Process steam 3 barg | 29 psig, preset, free from particles [kg/h] 7 15 15 15 15
Pure steam 2 barg | 29 psig, preset, free from particles [kg/h] 3 5 5 5 5
Coolant (supply line) 2 – 4 barg | 29 – 58 psig, [L/min] 5 5 5 5 5
preset (15°C) free from particles
Coolant (return line) 2 barg (29 psig) below supply [L/min] 5 5 5 5 5
Condensation Ambient pressure (max. temp. 98°C) [L/min] 1 1 1 1 1
Line voltage (TNS net): 208 VAC | 60Hz | 15 A, 400 VAC | 50Hz | 16 A
5-core cable: 3 + phase, 1 + ground, 1 + neutral universal-current-sensitive RCD 3 + 30 mA
Control unit Integrated: DCU controller, aeration system and up to 4 pumps
Control unit Industry PC
Housing material Stainless steel AISI 304
Display | Operation Touch panel 10“ | Touch screen
Interface with host PC Ethernet
External inputs
Scale connection maximum 2 RS 232
Analog inputs up to 3 (0 – 10 V)
External substrate pumps Up to 2; analog (0 – 10 V)
Aeration system
Microbial equipment O2 enrichment or gas flow ratio; maximum total flow rate: 1.5 vvm
Cell cultivatoin equipment Additive flow; maximum aeration rate: Overlay 1 vvm | sparger 0,1 vvm
Dual use Additive flow; maximum aeration rate: 1.5 vvm
Flow meter (rotameter) Air calibrated; 4 bara 20°C
Flow rates 0.6–60 mL/min up to 5–52 L/min
Accuracy +/– 4% FS
Thermal mass flow rate meter Air | N2, O2 or CO2
Flow range 0.6–30 smL/min up to 1–50 sLpm
Accuracy +/– 1% FS
Integrated pumps Up to 4 (2 + digital + 2 + digital | speed regulated)
Pump head – for silicon hoses with a wall Watson Marlow 102R
thickness of 1.6 mm | 1/16“
Available versions Controlled digitally (20 rpm) or speed regulated (5 – 50 rpm)
Flow rates Inner diameter of the hose: 0.5 mm 0.8 mm 1.6 mm 3.2 mm 4.8 mm
Flow rate: mL/revolution 0.02 0.05 0.22 0.81 1.66
Appendix 115
Supply unit Open piping frame
Material | surface roughnes Stainless steel AISI 316L | Ra < 0.8 μm (< 31.5 Ra)
(parts that come into contact with products)
Temperature control system Closed pressurized water temperature control system with recirculation pump, heat exchanger for cooling and
heating, optional electric heater
Operation (operation | sterilization): 8°C above cooling water temperature up to 90°C | 130°C
Heat exchanger (cooling | stainless steel) Stainless steel, copper-brazed | stainless steel, copper-brazed *option: Stainless steel welded
Electric heater (option) 5L | 10 – 30L 3 kW | 6 kW
Culture vessel 5L 10 L 15 L 20 L 30 L
H:D ratio 2:1 2:1 3:1 2:1 3:1 2:1 3:1 2:1 3:1
Total volume 6.8 L 15 L 15 L 22 L 22 L 30 L 30 L 42 L 42 L
Operating volume 5L 10 L 10 L 15 L 15 L 20 L 20 L 30 L 30 L
Minimum operating volume* 1.6 L 4.5 L 3.5 L 24 L 24 L 7.7 L 5.5 L 9L 7L
Weight of culture vessel cover and attachments
11 12 15 19 17 21 20 26 26
approx. [kg]
Permissible stirrer speed 20 – 20 – 20 – 20 – 20 – 20 – 20 – 20 – 20 –
1500 1500 1500 1000 1000 1000 1000 600 600
Engine output [kW] 0.5 0.8 0.8 0.8 0.8 1.2 1.2 1.2 1.2
Diameter of stirrer to culture vessel 0.4 0.4 0.4 0.4 0.4 0.4 0.4 0.4 0.4
[6-blade disk impeller]
Diameter of stirrer to culture vessel 0.5 0.5 N|A 0.5 N|A 0.5 N|A 0.5 N|A
3-blade segment impeller
Cover nozzles 1 + viewing window for illumination; not for 5 L
1 + nozzle for the exhaust cooler
1 + nozzle for the stirrer
1 + nozzle for the safety valve
4 + 19 mm nozzle (5 L and 10-3):
5 + 19 mm nozzle (10-2 – 30 L)
2 + handle
Upper nozzle level (not for 5 L) 3 + 25 mm nozzle
1 + nozzle for burst disk (only ASME culture vessels)
1 + lengthwise view glass
Lower nozzle level 4 + 25 mm port
1 + sensor nozzle for Pt100
Floor 1 + floor drain valve
Double wall 1 + supply
1 + return
Culture vessel design Double-walled stainless steel culture vessel with dished head and lengthwise view glass, stirrer from above 5 L:
Stainless steel | glass vessel
Material (in contact with product) Stainless steel AISI 316 L | borosilicate glass| EPDM (FDA)
Surface (vessel | attachments in contact with Ra ) 0.5 μm () 19.7 Ra) | Ra < 0.8 μm (< 31.5 Ra), electropolished
product)
Culture vessel design vessel | double wall 5L: -1 - +2.5 barg @ 150°C; 10-30L: -1 – +3 barg @ 150°C | -1 - +4 barg @ 150°C
Sensors | measuring range| readability
DO (“pO2”) Amperometrically or optically | 0–100% | 1% | 0.1%
pH Gel-filled | 2–12 | 0.01 pH
Foam | level | high foam Conductive, stainless-steel body with ceraminc isolation
Temperature culture vessel | temperature control Pt100 | 0–150°C | 0.1 C / Pt100 | 0–150°C | 0.1 C
system
pH | Redox Gel-filled | –1000 – 1000 mV | 1 mV
Pressure measurement Piezoresistive sensor | –0.5–2 [barü] | 1 mbar
Turbidity sensor One channel NIR absorption probe, gap width 10 mm or 20 mm | 0–6 AU | 0.01 AU
Standards CE | UL | CSA (EN61010, UL61010); culture vessel: ASME or PED or SELO (5L only PED)
116 Appendix
10.2 Connector pin assignment
X208 - ‘Serial-B’ (Option)
-X208 A120 / Port1

SUB-D Signal RJ 45
1 DCD (in) 1
2 RXD (in) 5
3 TXD (out) 4
4 DTR (out) 8
5 GND 6
6 NC
7 RTS (out) 2
PERLE RJ 45 / Male
M12 / Female 8 CTS (in) 7
X209 - ‘Ext. Signals-A/B’
Pin Signal Tag

1 AI 01 EXT-A
2 AI 01 GND EXT-A
3 AI 02 EXT-B
4 AI 02 GND EXT-B
5 NC
6 NC
7 24VDC/F04
M12 / Female 8 GND24V
X210 - ‘Ext. Signals-C’
Pin Signal Tag

1 AI 03 EXT-C
2 AI 03 GND EXT-C
3 NC
4 NC
5 DO 21A COMAL
6 DO 21B COMAL
7 24VDC/F04
M12 / Female 8 GND24V
X211 - ‘PUMP-C’
Pin Signal Tag

1 NC
2 NC
3 GND AO 02 GND PUMP-C
4 AO 02 PUMP-C
5 NC
M12 / Female
Appendix 117
X212 - ‘PUMP-D’
Pin Signal Tag

1 NC
2 NC
3 GND AO 03 GND PUMP-D
4 AO 03 PUMP-D
5 NC
M12 / Female
10.3 EC Declaration of With the attached declaration of conformity, Sartorius Stedim Systems GmbH
Conformity confirms the compliance of the BIOSTAT ® Cplus bioreactor with the directives cited.
The signatures on the English edition are representative for the Declarations of
Conformity executed in the other languages.
118 Appendix
Appendix 119
10.4 Dimensioning the Variable The measuring jets of the variable area flowmeters are designed to match the gases,
Area Flowmeters e.g. for air or nitrogen. If you use flow rate meters for gases they are not designed for,
this can produce gas flow rates that are too large or too small.
− Flow rate meters are normally calibrated and scaled for standard conditions. The
information can be found on the glass test tube or holder. Standard calibration
conditions are e.g.:
− Gas type: Air
− Temperature: 20 °C = 293 K
− Pressure: 1 bar overpressure
t Check the variable area flowmeters that your bioreactor is equipped with to see
which gases it has been calibrated for under which conditions.
If the exact flow rates of a gas must be known for the evaluation of a process and
other operating conditions are available, like the calibration (e.g. other gases at other
pressures and temperatures), the measured flow rates must be converted for the
respective gas.
The manufacturer of variable area flowmeters can furnish documents that allow the
flow rates to be determined for certain gases under defined operating conditions and
|or suitable correction factors to be determined for measured flow rates.
10.5 Decontamination When returning equipment, copy the following form as required, carefully complete
Declaration it and enclose it with the delivery documents.
The recipient must be able to inspect the completed declaration before removing the
device from the packaging.
120 Appendix
Decontamination Declaration
Declaration Concerning the Decontamination and Cleaning of Equipment and Components

To protect our personnel, we require that all equipment or components which come into contact with our personnel
at customers’ facilities be free of biological, chemical, or radioactive contaminants. We will only accept an order
when:
– the equipment or components have been adequately CLEANED and DECONTAMINATED.
– this declaring document has been completed, signed and returned to us by an authorized person.
Please help us in assuring a safe, hazard-free work environment.
Description of the Equipment or Component(s)

Description | Cat. No.:
Serial no.
No. of invoice | Delivery note:
Date of delivery:
Contamination | Cleaning
Attention: Please specify exactly the biological, Attention: Please describe the cleaning and
chemical, or radioactive contaminant decontamination procedure | method.
The equipment was contaminated with: and it has been cleaned and decontaminated by
Legally binding declaration

I (we) certify that all information given in this form is correct and complete. The equipment and components have
been adequately decontaminated and cleaned according to the legal requirements. No chemical or biological or
radioactive risks remain that could endanger exposed persons’ safety or health.
Company | Institute:
Address | country:
Tel.: Fax:
Name of authorized person:
Position:
Date | Signature
Please pack the equipment properly and send it

Sartorius Stedim Systems GmbH
ex recipient to your local service representative
Service Department
or directly to Sartorius Stedim Biotech GmbH.
Robert-Bosch-Str. 5–7
34302 Guxhagen
Germany
© 2012 Sartorius Stedim Biotech GmbH
Appendix 121
122
Part B: DCU4 System for BIOSTAT® Cplus
Operating Manual
(Original Operating Manual)
Digital Measurement and Control Unit
123
11. User Information
11. User Information
This operating manual shows the standard functions of the DCU software.
DCU systems can be customized according to the customer's specifications.
Therefore, this documentation may describe functions that a delivered configuration

does not contain, or a system may contain functions that are not described here.
Information about the actual scope of functionality can be found in the configura-
tion documents. Additional functions can be described in the technical data sheet in
the overall documentation.
Illustrations, parameters, and settings in this documentation are only examples. They
do not show the configuration or operation of a DCU system in terms of a particular
unit, unless they explicitly refer to that unit. Information about the exact settings are
provided in the configuration documents or have to be obtained empirically.
Usage Instructions, Structure, and Functions

The DCU system can be integrated into higher-level automation systems. For example,
the industrially tested MFCS system can take on host PC functions like process
visualization, data storage, process logging etc.
This operating manual shows operating values and settings that are default values
and examples. Only if explicitly specified as such will they show settings for the
operation of a particular bioreactor. Information about the settings that are
permitted for a bioreactor and the specifications for a customer system can be
found in the configuration documentation.
Only system administrators or authorized, trained, and experienced users may change
the system configuration.
124 User Information

12. System Behavior
During Start 12. System Behavior During Start
The control unit is turned on together with the whole system using the main switch.
After switching on the system and program start (or when the power comes back on
after a power outage), the system starts in a defined basic state:
− The system configuration is loaded.
− Any user-defined parameters from a previous process are stored in a battery-
buffered memory and can be used for the next process:
− Setpoints
− Calibration parameters
− Profiles (if there are any)
− All regulators are turned off, and actuators (pumps, valves) are in the rest position.
System Behavior During Start 125

For interruptions in operation, the startup behavior of outputs and system functions
that have a direct effect on the associated end unit (regulators, timers, etc.) depend
on the type and duration of the interruption. There are several different types of
interruptions:
− Turning the unit off | on at the main switch of the control unit.
− Failure of power supplied from the connection in the laboratory (power outage).
In the “System Parameters” submenu of the main “Settings” menu, a maximum

duration for power interruptions can be set as “Failtime”:
In case of a power outage shorter than “Fail Time”, the system continues as follows:
− A “Power failure” error message shows the outage time and duration.
− Controllers continue to work with the set target value.
− Timer and target value profiles continue to be processed.
If the power outage lasts longer than the configured “Failtime”, the DCU system acts
as though the user had turned the unit off normally, that is, it starts in the defined
basic state.
After the next restart, the alarm message “Pwf stop ferm” message [¨ alarm
messages in appendix “20. Appendix”], specifying the date and time at which the
power failure occurred.
126 System Behavior During Start

13. Principles of Operation
13. Principles of Operation
13.1 “Main” Menu The “Main” menu offers a graphical overview of the controlled unit, with symbols
for the reactor, components of the gas supply (e.g. valves, MFCs), sensors, pumps,
dispensing counters, and if applicable, additional peripherals with their typical
locations relative to the reactor.
“Main” Menu operator screen
Header, display of the system status and the active menu

Work area, display of the function elements:
− Gas feeds air, O2, N2, CO2, e.g. with access to associated submenus
− Intake of correction materials ACIDT, BASET, AFOAMT, LEVELT and with dispenser
counters and pumps
− Measured value display and access to submenu for configuration of STIRR, LEVEL,
TEMP, pH, DO (“pO2”) by pressing the function elements (implemented as touch
keys)
Footer with function keys for:

− Access to the menus for associated main functions
− Activation of additional functions such as
− Remote operation (external computer)
− “Alarm” menu with overview of alarms
− Emergency off (”shutdown”)
Principles of Operation 127

13.1.1 Work Area − The measuring range shows the function elements and submenus of the active
main function:
− preselected process values with current measured or set value
− pumps or dispenser counters with process values, e.g. flow rates or dispensing
volumes for correction materials and gases
− controllers, e.g. for temperature, speed, mass flow controller (MFC) etc.,
with current setpoints
− sensors, e.g. for pH, DO (“pO2”), antifoam, etc., with measured values
− peripheral devices, e.g. weighing system, with measured values or current
setpoints
“Controller” Menu operator screen
− The DCU system is operated directly on the display by selecting a main function
and the associated submenus. The function elements in the work area and the
menu buttons in the footer contain touch keys. By pressing them, you can activate
the associated submenus, such as for the input of data and set values or the
selection of operating modes.
− Available functions, tag names, parameters, and submenus depend on the
controlled device for which the DCU system is intended and on the configuration.
128 Principles of Operation

Reference Time Input Field for Process Start
By pressing the input field in the “Main” menu for the corresponding subsystem (see
illustration below), you can enter a reference time for a process or set the time to
zero (time in format [hh:mm:ss]).
− Only available in selected configurations
In the work area, the reference time is only displayed in the [hh:mm] format.
The complete format [hh:mm:ss] can be viewed in the submenu for input of the
reference time.
13.1.2 Header The header of the screen only shows status information:
2009-05-13 14:55:09:
Date in format [yyyy-mm-dd]; time in format [hh:mm:ss]
All alarm messages are shown in the “Alarm” menu.

Alarm display:
− Bell white: no alarm
− Bell red: Alarm has occurred, information about alarm in alarm message [¨ list of
alarm messages in the “20. Appendix” chapter] and in “Alarm” menu.
13.1.3 Footer The footer includes the main function keys for switching between main functions:
Display:
− Selected main function: Button light gray, activated
− Function not selected: Button dark gray, deactivated

13.2 Display of Function
Elements The display of the function elements in the working area shows their current status
and the intended use:
Symbol Display Meaning, use

Function element [PV tag]: Field for short name (”tag”) for the function element, e.g. TEMP,
Button with gray STIRR, pH, DO, ACID, SUBST_A, VWEIGH
underline MV [Unit]: Field for measured or set value in its physical units
Submenu or function can be selected by pressing
Function element Measured value collection or output of function element is active,
Button with green with measured value or set value as shown
underline
Function element Output of function element is active, controller in cascade mode
Button with light
green underline
Function element Display of function if in “Manual” mode; (switched on or off);
Button with yellow automatic control not possible
underline
[Tag PV] No underline No submenu assigned (function cannot be selected)
MV [Unit]
“U”, “V”, “Y”, “Z” Arrow button Move forward or back in specified menu or function
Pump off Æ Direct access to submenu to select the mode
Auto on
Æ Line gray Æ green
Pump off Æ − Submenu to select operating mode:
Manual on [¨ Example in Chapter “14. “Main” Menu”]
Æ Yellow underline,
pump gray Æ
green
Valve off Æ Direct access to submenu to select mode, example for 2/2-way valve
Auto on
È Line gray Æ green
Valve off Æ Valve symbol also shows flow direction (possibly changed)
Manual on − Submenu to select the operating mode:
È Yello underline, [¨ Example in Chapter “14. “Main” Menu”]
flow direction
green
− Examples for function elements, short descriptions, measured values,

working values, and submenus that can be called by selecting touch keys
[¨ Chapter “14. “Main” Menu”, Section “14.3 Direct Access to Submenus”].

13.3 Overview of the
Menu Buttons
Button, key, symbol Meaning, use

“Main” menu button Start screen with graphical overview of the controlled unit:
− Display of components of the current configuration
− Overview of measured values and process parameters
− Direct access to important menus for operating input
“Trend” menu button Display of process sequences, selection of 6 parameters from:
− Process values
− Setpoints from control loops
− Outputs of controllers
“Calibration” menu button Menus for calibration functions, for example for:
− Measurement sensors for pH, DO
− Totalizer for all pumps (ACID, etc.)
− Totalizer for aeration rates for valves
− Scales
“Controller” menu button Operating and configuration menus for controllers, for example:
− Temperature regulation TEMP
− Speed regulation STIRR
− pH control and DO control
− Control of correction medium pumps (e.g. pH, FEED)
− Aeration rate regulation (valves, or mass flow controllers).
“Phases” menu button Function for programmed sequences (time or step controls), for example:
− Sterilization
− Pressure hold test of culture vessel
“Settings” menu button Basic system settings, for example:

− Measurement ranges of process values
− Manual operation, e.g. for inputs and outputs, controllers, etc.
− External communication (e.g. with printers, external computers)
− Selection, modification of configurations (password-protected, only for authorized service
technicians)
“Remote” menu button Operation with external computer systems (central computer):
− Pressing the menu button switches to remote operation; instructions on configuration
[¨ Chapter “19. “Settings” Menu”]
“Alarm” menu button Overview table of alarms that have occurred:

− If alarms occur, the symbol changes color and an acoustic signal sounds.
− Display red : Table still contains unconfirmed alarms
− Pressing the menu button opens an overview menu of all alarm messages.
“Shutdown” menu button Emergency stop function:
− Pressing the menu button puts all analog and digital outputs into a safe state (red color
change on the button indicates shutdown activated).
− Repeated pressing of the button cancels the shutdown state and restores the original state.
Main functions can be selected at any time during a running process. The title of the
main function shown in the work area is also displayed in the header.

13.4 Overview of Selection Keys
Cancel
− Changes will not be saved
Confirmation of input
Additional controller functions
Additional phase parameters
Cancel
− Changes will not be saved
Deleting characters
Selection of sign when entering a value
Selection list of process values
13.5 Direct Function Keys for The function elements in the work area of the “Main” menu can contain function keys
Selection of Submenus that can be used to directly active submenus for important functions:
− for the numerical input of setpoints, conveying and flow rates etc.
− for the configuration of alarm limits
− for the selection of controller modes
Which functions can be reached from the main menu depends on the configuration.
Press the function keys to view the available functions in the supplied configuration.
The “Direct Access to Submenus” section in the chapter on the “Main” menu shows
examples of the screens and submenus reachable from the direct function keys.
Detailed instructions for the associated functions and possible inputs can be found
in [¨ Chapters “16. “Calibration” Menu” and “17. “Controller” Menu”.
Example: Input of Temperature Setpoint:

t In the work area of the “Main” menu, press the TEMP function element or in the
work area of the “Controller” menu select the TEMP controller (TEMP function
element).
− When the “Main” menu is accessed, a submenu opens with a keypad on the
left side for data entry and a selection field for possible operating modes
[¨ Fig. 13-1].
− When the “Controller” menu is accessed, the “Setpoint” touch key can be used
to enter a set value (after pressing the touch key, a screen keyboard also opens).
The touch key (1) can be used to select the mode [¨ Fig. 13-2].

Fig. 13-1: Setpoint input and selection of the “TEMP” controller mode from the “Main” menu
Fig. 13-2: Setpoint input and selection of the “TEMP” controller mode from the “Controller” menu
t Enter the new setpoint using the screen keyboard (note the permissible value
range under the input field). If you want to correct the value entered, press the
BS key. If you don't want to save the new value, leave the submenu by pressing
the C key.
t Confirm by pressing the “OK” key.
y The submenu window closes.
The setpoint is active and is displayed.

Example: Selecting the Controller Mode:
t In the work area of the main menu, press the TEMP function element or select the
“Controller” function and there the TEMP controller.
t Press the function key for the desired mode on the right side.
t Confirm by pressing the “OK” key. The function (of the controller) is activated and
displayed.
You can access the full operator screen of the controller through .
This corresponds to activating the “Controller” menu button
and selecting the TEMP controller there in the overview screen
[¨ Chapter “17. “Controller” Menu”].
13.6 Selection Lists and Tables If submenus contain lists of elements, short names, or parameters that cannot be
displayed in a window, a scrollbar with a position marker is displayed.
Trend display operator screen submenus
To page through lists that contain more entries than can be displayed in the window:
t Press the arrow key “V” (down) or “U” (up).
t Press the position mark (light gray field in the scrollbar) and push it.
Press directly in the scrollbar at the relative height where the channel tag could be
located.

14. “Main” Menu
14. “Main” Menu
14.1 General Information The “Main” menu opens after the control unit has been switched on.
This is the central starting point for in-process operation.
“Main” Menu start screen
The graphical display of the system structure simplifies the overview of the system
components and uses function elements implemented as touch keys to provide access
to the submenus for the most important or most frequently used settings. If practical,
the function elements also show the currently entered or configured measurement
values and settings.
Which function elements are actually shown depends on the configuration of

the DCU system, the unit controlled, e.g. the type of bioreactor, and customer
specifications.
14.2 Process Displays in the The function elements can display associated process values:
“Main” Menu − Values measured by connected sensors such as pH, DO (“pO2”), foam, etc.
− Calculated variables like pump filling amounts, calculated values of arithmetic
functions, etc.
− Process duration displays
− Measured data and key figures from the responses of external components such as
speed regulation, mass flow controllers, scales, etc.
“Main” Menu 135

14.3 Direct Access to The following menu screens show examples of submenus accessible from the “Main”
Submenus menu and configuration options for the measurement and control system. Which
submenus are available and which parameters can be configured depends on the
configuration:
Target value specifications and mode selection for headspace aeration (overlay) for
air and CO2 and media aeration (sparger) for all gases; example menu “O2 SP”
Settings for alarm limits and activation of alarm monitoring for totalizer, example
“BASET”
Agitator speed “STIRR”
Mode selection for foam monitoring “AFOAM”,

Analogous for level control “LEVEL”
136 “Main” Menu

15. “Trend” Menu
15. “Trend” Menu
15.1 “Trend” Display Using the “Trend” display, the user can graphically display process values for a period
of up to 72 hours. This overview of the process flow gives for example a quick
impression as to whether the process is running as expected or whether irregularities
or disruptions are present. The trend display works retroactively from the current
point in time and offers:
− Up to 8 (selectable) channels
− Time basis 1 – 72 hours
Start screen for the “Trend” menu
Field Value Function, entry required

Key line 1…8 Display and set channels
Diagram 1…8 Line chart of the selected channels (y) over time (x)
Top Upper limits of the selected display ranges for each
channel
Center Colored line chart
Bottom Lower limits of the selected display ranges for each
channel
Subtitle HH:MM:SS Time scale
“Trend” Menu 137

15.2 Configuring the “Trend”
Display
15.2.1 Setting the Trend

Display for Parameters
t Select the “Trend” menu button.

t In the header, press the key of the channel that you want to set:
y the “Channel # Settings” window opens.
t To change the parameter for the channel, press “PV”.

y The “Select Buffered Channel” menu shows the default values:
t Press “Cfg” to display all parameters in the configuration. If the desired parameter
is not visible, you can scroll through the table.
t Press the key for the parameter to select it.
y The parameter will be activated immediately.
t To deselect a parameter without having to re-assign the channel, press “.....”.
15.2.2 Setting the Parameter

Display Range
t Select the “Channel # Settings” window and press “Min” and/or “Max”.
t Enter the upper and | or lower limit.
y The display limit values for the parameter are shown underneath the data window.
t Confirm the input with “OK”.
138 “Trend” Menu

15.2.3 Resetting the
Display Range
t Press “Reset Range” on the “Channel # Settings” screen to reset a modified display
range back to the factory setting for “Max” and “Min”.
15.2.4 Setting the Trend The color for every parameter can be selected from a table.
Display Color: t Select the “Channel # Settings” screen and press the key with the name of the
preselected color.
t Press the key with the name of the new color to be used.
The selection is instantly assigned and activated.
15.2.5 Defining a New Time

Range as “Time Range”
t Enter the value for the time range (e.g. 1 h).

y The time scale below in the working environment changes automatically.
y The parameter trend will be displayed over the new time range.
“Trend” Menu 139

16. “Calibration” Menu
16. “Calibration” Menu
16.1 General Information In the “Calibration” menu, all calibration actions required for routine operation can
be activated:
− Calibration routines for sensors: e.g. pH, DO (“pO2”), turbidity
− Sensor function tests: e.g. Redox
− Calibration of pump filling counter: e.g. acid, base, substrate
− Calibration of gas filling counter: e.g. N2, O2, CO2
Start screen for the “Calibration” menu
After pressing the “Calibration” menu button, the Calibration menu opens. Selectable
touch keys show the status of the associated calibration functions and open the
associated submenu to carry out calibration routines.
Operating instructions on the individual steps and required entries on the display lead
the user through the menus.
The calibration parameters remain stored when the DCU system is switched off.
After power is restored, the DCU system uses the saved figures until a new calibration
is carried out.
140 “Calibration” Menu

16.2 pH Calibration Conventional pH sensors are calibrated using a two-point calibration with buffer
solutions. During pH measurement, the system calculates the pH value based on the
sensor potential according to the Nernst equation while taking zero point deviation,
slope and temperature into consideration.
During calibration you can enter the reference temperature manually; during pH
measurement temperature compensation is carried out automatically based on the
temperature measurement value in the bioreactor.
Sensors are calibrated before installation at the point of measurement, e.g. in the
culture vessel. Sterilization can alter the sensor zero point. To recalibrate the pH
sensors, you can measure the pH value externally in a sample taken from the process
and enter the value into the calibration menu. The calibration function compares
the pH value measured online with the one determined externally, calculates the
resulting zero offset and displays the corrected process value.
The effects of heat during sterilization and reactions of the diaphragm and/or
electrolytes with components of the medium can influence the measurement
properties of the pH sensors. Test and calibrate the pH sensors before each use.
The operator screen for the pH sensors shows both the pH value and the
measurement chain voltage of the sensors, as well as the zero offset (”zero”) and
slope sensor parameters. That allows you to easily check the functionality of the
pH sensors.
16.2.1 Calibration Sequence t Press the touch key of the sensor to be calibrated (”pH-Measure”) in the
“Calibration” menu.
t Start calibration by pressing “Measure”.
t Select the desired calibration function.
Touch keys:
− “Calibrate”: Complete calibration cycle with zero point calibration “Zero” and slope
calibration “Slope”.
− “Recalibrate”: Recalibration [¨ Section “16.2.2 Recalibration”]
− “Calibrate Zero”: Zero Point Calibration
− “Calibrate Slope”: Slope Calibration
“Calibration” Menu 141

t Select the type of temperature compensation
If “Manual” is selected, the following input window opens:

If “Auto” is selected, the menus below open immediately:
t In the “Zero Buffer” submenu, enter the pH value to calibrate.

Confirm the value entered with “OK”:
t Observe the measured value display in the “Zero Value” submenu.

Once the display is stable, confirm the measurement with “OK”:

t In the “Slope Buffer” submenu, enter the pH value to calibrate.
Confirm the value entered with “OK”:
t Observe the measured value display in the “Slope Value” submenu.

Once the display is stable, confirm the measurement with “OK”:

Mode Measuring, Calibrating, Recalibrating
pH pH pH value display and | or entering the pH of the
external sample during recalibration
Electrode mV Combination electrode voltage (raw signal)
TEMP °C Temperature value for temperature compensation
Zero mV Display the zero offset
Slope mV/pH Display the slope
Measure Automatic switch to pH measurement after calibration
routine is completed
Calibrate Starting the calibration routine
Recalibrate Starting the recalibration
Calibrate Zero Zero point calibration as a single step
Calibrate Slope Slope calibration as a single step
Manual Manual temperature compensation with entry of a
value measured outside the culture vessel
Auto Automatic temperature compensation with the value
measured inside the culture vessel

16.2.2 Recalibration By following the operating steps described below, you can adapt the calibration of
a pH sensor to changed measuring conditions after a sterilization cycle or during the
process as needed:
t Measure the pH value of a current sample in-process.
Use a precise and carefully calibrated measurement device.
t Press the touch key of the pH sensor to be calibrated.

t Press the “Measure” touch key and select the desired calibration.
t For recalibration, press “Recalibrate” and enter the pH value measured externally
in a sample:
y The DCU system calculates the zero offset and displays the corrected pH value.
16.2.3 Special Notes − Whenever possible, use buffer solutions manufactured by the sensor manufacturer
as contained in the equipment supplied with the pH sensor. Information on
reordering is available on request.
− If the “zero offset” and “slope” values are known and the process allows, you can
also enter these values directly into the relevant fields.
− The sensor's service life is limited and depends on the in-process working and
operating conditions. Whenever a function check or calibration points to a
malfunction, the pH sensor should be serviced and replaced as needed.

− The pH sensors must be serviced or replaced when the following values are outside
the specified range:1
− Zero point offset (”zero”) outside -30 to +30 mV
− Depending on the type and design of the sensors supplied, the menus, sequence
and operation of the calibration function can differ from the information
provided herein. Please refer to the configuration documents or to the function
specifications of the bioreactor, if available.
16.3 DO Calibration Calibration of the DO electrode is based on a two-point calibration. Calibration is

performed in [% oxygen saturation]. Calibration determines the sensor parameters
zero current (”zero”) and slope. The reference value for “zero” is the oxygen-free
medium in the culture vessel. Air-saturated medium can be defined as 100%
saturated and be the basis for determination of the slope. Since you will be
calibrating the sensor after sterilization, take changes in the measuring properties
that can result from heat exposure or effects of the medium during sterilization into
consideration.
The operator screen for calibrating the DO sensor corresponds to that of the
pH calibration. Follow the description for pH calibration [¨ Section “16.2 pH
Calibration”] of this manual or the operator screen for DO calibration on your DCU
system. In addition to DO saturation, the operator screen also shows the current
sensor current as well as the zero current and slope with calibration conditions.
This allows easy regulation of the sensor's functions.
1 Limit values may differ depending on the construction and manufacturer of the pH sensors;
consult the manufacturer's documentation.

16.3.1 Calibration Sequence t Press the touch key of the sensor to be calibrated (”pO2 Measure”) in the
“Calibration” menu.
Selection of Operator Screen for the DO (“pO2”) Sensor

16.3.1.1 Zero Point
After performance of in-situ sterilization, do not yet aerate the culture vessel with air
Calibration
or the intended oxygen-containing gas.
Before starting zero point calibration:

t For an exact zero point calibration, aerate with nitrogen until the oxygen dissolved
in the medium is displaced.
t Start calibration by pressing “Measure”:
t Select the desired calibration function.
Touch keys:
− “Calibrate”: Complete calibration cycle with zero point calibration “Zero” and slope
calibration “Slope”.
− “Calibrate Zero”: Zero Point Calibration
− “Calibrate Slope”: Slope Calibration
t Select the type of temperature compensation

y If “Manual” is selected, the following input window opens for the temperature.
y If “Auto” is selected, the menus below open immediately:
t In the “Zero Buffer” submenu, enter the value to calibrate for oxygen saturation,
in percent. Confirm the value entered with “OK”:
t Observe the measured value display in the “Zero Value” submenu. Once the display
for the DO (“pO2”) value near 0 % is stable and shows a zero current in the range
from 0 to 10 nA, confirm the measurement with “OK”.

16.3.1.2 Slope Calibration t Adjust the agitator speed, temperature, and if necessary the pressure for
the process using the appropriate controllers [¨ Chapter “17. “Controller” Menu”].
Aerate the culture medium with the planned gas mixture or e.g. air until oxygen
saturation has been achieved.
t Start the calibration function as described in section 16.3.1.1. In the “Calibration
Mode” submenu, select the “Calibrate Slope” calibration function.
t Select the type of temperature compensation.
y If “Manual” is selected, the following input window opens for the temperature.
y If “Auto” is selected, the menus below open immediately.
t In the “Slope Buffer” submenu, confirm the value to calibrate for oxygen
saturation in percent with “OK”.

t Observe the measured value display in the “Slope Value” submenu.
t Once the measured value for the sensor current is stable near 60 nA, calibrate the
slope by confirming with “OK”:
16.3.2 Special Notes Prior to first use or whenever the DO (“pO2”) sensor has been disconnected from the
power supply (measurement amplifier) for longer than 5 to 10 min., it has to be
polarized. Polarization can take up to 6 hours (less time when the sensor was only
disconnected from the measurement amplifier for a few minutes). This does not apply
to optical DO sensors.
Follow the sensor manufacturer's instructions.
If necessary, you can enter the zero offset and slope directly into the relevant
submenus:

The DO (“pO2”) sensor must be serviced if:
− The zero point (”Zero Value” submenu) is not within the range of 0 .. +10 nA,
− The sensor current at maximum aeration with air (”Slope Value” submenu) is lower
than 30 nA.
16.4 Calibrating the The turbidity sensor works on the principle of light absorption and measures the
Turbidity Sensor turbidity of liquids.
Calibration of the turbidity sensor determines the sensor zero point using a single-
point calibration.
The DCU system calculates the turbidity in absorption units (AU) from the zero point
deviation. It calculates the average of measurements over a defined time window, the
damping factor DAMP. To obtain stable process values, you can select DAMP in 4
setting ranges.
The operator screen for the turbidity sensor shows both the absorption units (AU) as
well as the raw signal from the sensor in [%], along with the deviation from the zero
point for “0 AU”. This allows you to easily check the turbidity sensor's functional
capacity.
Turbidity sensor calibration operator screen

Field Function, entry required
Mode Mode key, selection of “Measure” | “Calibrate”
Turbidity Display the process value in [AU]
Electrode Display the sensor raw signal in [%]
Zero Display the zero point after calibration in [%]
Damping Set and display signal damping: 6 s, 12 s, 30 s, 60 s
16.4.1 Calibration Sequence t Hold the sensor in the “zero point solution”.
t Select the “Calibration” function and press the touch key for the turbidity sensor,
“TURB Measure”.
t In the “Calibration TURB” menu, press the “Measure” mode key.
t Select the “Calibrate” touch key in the submenu.
Submenu is closed again after key pressed. Mode switches back to “Measure” after
key pressed.
16.4.2 Special Notes Depending on the process requirements, you can calibrate the light absorption before
inoculation and aeration as a reference parameter. This is possible in particulate- and
bubble-free water in a suitable buffer, deionized water or in the culture medium
directly in the culture vessel.

16.5 Redox Calibration The Redox calibration covers a function test of the Redox sensor (measurement of the
Redox value of a reference buffer).
During sterilization, thermal effects and interactions with the culture medium can
impair the measurement properties of the Redox sensor.
You should therefore always check the sensor prior to use.
Redox sensor calibration operator screen

REDOX mV Display the combination electrode voltage measured in the
reference buffer
Electrode mV Combination electrode voltage of the last calibration
Check mV Input of a reference current of the reference buffer for the
Buffer current temperature of the reference buffer (information on
the buffer bottle)

16.5.1 Functional Test The functional test of the Redox sensor is carried out prior to installation in the
culture vessel, i.e., before sterilization.
t Fill the reference buffer into a graduated beaker and place the Redox sensor in it.
t Select the “Calibration” function and press the touch key for the Redox sensor.
t Press “Check Buffer” and enter the reference value of the buffer in [mV], as
indicated for the current temperature on the buffer bottle.
16.5.2 Special Notes When there are deviations of more than 6 mV (approx. 3 %), the Redox sensor must
be serviced. Follow the manufacturer’s instructions on the documents supplied with
the sensor.

16.6 Totalizer for Pumps For documenting the correction medium consumption, the DCU system adds up the
and Valves running times of the pumps or proportioning valves. It calculates the feed volumes
from the running times, taking the specific flow rates into account. You can
determine unknown pump feed rates using the calibration menus of the pumps or
proportioning valves and you can enter known specific feed rates directly into the
calibration menus.
The calibration and filling counter functions are the same for all pumps and
proportioning valves. Therefore, this section only describes the calibration for one
of the “ACIDT” acid pumps.
Operator Screens

Operator Screens

Mode Calibrate Start the “Calibrate” or “Reset” routine:
Totalize − After completion of “Calibrate”, the system automatically
Reset switches to “Totalize”
− Reset sets the filling counters back to zero
ACIDT ml Display the volume of liquid being pumped:
− BASET etc., for the alkaline pump
− AFOAMT for antifoam pump
− LEVELT for level pump
Flow ml/min Enter the specific feed rate of the pump or the flow rate of
the proportioning valve, if known

16.6.1 Pump Calibration
Always use the same type of tubing with the same dimensions for calibrating and
Sequence
pumping the media.
t Place the tubing end of the pump inlet in a beaker filled with water and the tubing
end of the pump output in a measuring beaker to measure the feed volume in
liters.
t First, fill the tubing completely with the medium. For this purpose, you can switch
on the pump manually.
Pump Calibration Operator Screen

t Press the touch key of the pump to be calibrated.
t Select the touch key for the operating mode. Prior to the first calibration,
it displays the “Off” operating mode.
After a calibration has run, it switches to “Totalize”.
t Select the “Calibrate” touch key in the “Mode” submenu.
Fig. 16-1: Operating Type Selection
y The “START calibration with OK” menu is displayed.

t Start the pump calibration with “OK”. The “STOP calibration with OK” menu is
displayed. The pump pumps the medium.
t Once a sufficient volume has been transferred, press [“OK”].
t On the graduated beaker, read off the feed volume and enter it in the “ACIDx_T:
Volume“ submenu.
y The DCU system calculates the pumping rate automatically from the internally
registered pump run time and the pumping quantity calculated.
y The pumping rate is displayed in the “Calibration ACIDT” submenu in the “Flow”
field.
Activating the Filling Counter

The filling counter is activated after the calibration routine is completed and the
related controller is automatically switched on.
Special Notes
If you know the feed rate of the pump, you can directly enter this rate after pressing
the “Flow” input field.
t Press the “Flow” touch key.

t Enter the appropriate value on the keyboard.
t Confirm the value and start the calibration with “OK”.
You can set the fill counter to zero using the calibration function
[¨Fig. 16-1, “Reset” Mode].
16.6.2 Scale/Balance The weight of bioreactors (culture vessels), feed bottles or media or harvest containers
Calibration Sequence can be measured on weighing platforms or pressure gauges.
Any tare corrections required, e.g. after re-equipping the culture vessel or refilling a
holding bottle, can be made during running operations. To do so, determine the net
weight and adapt the tare weight to the change in weight caused by the changed
equipment.

Scale Calibration Operator Screen

xWEIGHT/ g/kg Display net weight (WEIGHT = gross tare)
FEEDW-x# − WEIGHT: weight of culture vessel
Tare g/kg Display tare weight
Gross g/kg Display gross weight

Example of Culture Vessel Calibration
t On the operator screen, press the “VWEIGHT Measure” touch key.
t Press the “Mode” touch key and select “Tare” (1) for zero taring.
t Press the “Mode” touch key and select “Hold” (2) to detect weight changes.
t Read the measured weight change off and end the measurement with “OK”.
t In the “Calibration VWEIGHT” submenu, enter the weight change into the “Tare”
field using the screen keyboard.
t Confirm the input with “OK”.

17. “Controller” Menu
17. “Controller” Menu
17.1 Functional Principle The control loops in the DCU system work as PID controllers, setpoint generators or
and Equipment two-point controllers and are adapted to their control circuits. PID controllers can be
parameterized to match the control task. The controller outputs control their control
elements either continuously or using pulse-width modulation. Single-sided and split
range control are implemented.
Which controllers are implemented in a DCU system depends e.g. on the device
(e.g. bioreactor). Controllers can be customized.
Available controllers in the DCU software include:
Controllers Function
“TEMP” temperature controller PID cascade controller with pulse-width modulated split range outputs for the
control of the heater and | or valve on the cooling water intake with the measured
value of the culture vessel temperature as controlling value
“JTEMP” double wall temperature Slave controller for temperature control:
controller − with TEMP controller “off”, possible as setpoint generator for heating
Speed regulation “STIRR” Setpoint generator for external motor controller controlling the agitator motor
pH controller “pH” PID controller with pulse-width modulated split range outputs:
− controls the acid pump and | or the CO2 supply and the alkaline pump
DO controller “pO2” PID cascade controller for controlling up to 4 slave controllers:
− Gas filling controller Air, O2 or N2
− Gas Flow Controller
− Speed regulator
− Controller for substrate supply
Gas Filler Controller Slave controller or setpoint generator for gas proportioning valves, pulsed feed:
AirOv, AirSp − Air for headspace (overlay) and medium aeration (sparger)
− O2 for medium aeration
O2
− N2 for medium aeration
N2 − CO2 for headspace (overlay) and medium aeration (sparger)
CO2
Gas Flow Controller Slave controller or setpoint generator for mass flow controller
− Each of the gases listed in each segment
Antifoam controller “FOAM” Pulse pause controller for introduction of antifoam agent “AFOAM”
Level controller “LEVEL” Pulse pause controller for level controller “LEVEL”
Substrate controller “SUBSA/B” Target value generator for filling pumps
Weight controller PID controller with pulse-width modulated output for harvesting pump;
works with the weight of the culture vessel “VWEIGHT” as master variable
Gravimetric filling control “FLOW” Setpoint generator for internal or external filling pump; works with the weight of
the substrate vessels “BWEIGHT”, “FWEIGHT” as master variable:
− Only controlled devices with associated weight measurement
Pressure controller “PRESS” PID controller with constant output for pressure control valve:
− Only controlled devices with pressure regulation
The “Profile Parameter” function can be used to navigate to the setpoints of the
individual controllers. Time-based setpoint profiles can be set up.
Up to 15 steps can be configured.
Any DCU system pre-installed at the customer's facilities can be additionally

retrofitted with control functions by changing the configuration. Moreover, control
blocks available in the software can also be used to configure sensor controllers.
164 “Controller” Menu

Switching between controller operating modes is mostly shock-proof:
off Controller switched off with defined output
auto Controller activated
manual Manual access to control element
profile Selection of previously defined profile. If no profile is defined,
the controller automatically switches to “auto” mode
In the controller operator screen you can enter the nominal value, operating mode
and controller output. The control ranges depend on the configuration. With a
password, you have access to the parameterization screen to set PID parameters,
output limits, and if necessary a dead zone. In “Remote” operation, the host PC
defines the setpoints and operating modes.
17.2 Controller Selection You can access the operator screens on the controllers of a configuration in various
ways:
− For the controllers most frequently used, from the “Main” menu or from the
“Controller” menu.
17.3 General Controller For the most part, operation of the controller is uniform. It comprises setting the
Operation setpoints and alarm limits and the selection of the control operating mode. If a
controller can control more than one output, the controller output is assigned by
means of the parameterization functions accessible with a password. This also
applies to controller settings not required during routine operation.
“Controller” Menu 165

Operator Screen
Fig. 17-1: Selecting the Temperature Controller
Field Display Function, entry required

Controller Selection Input of the controller mode
mode
Function key off Controller and slave controller switched off
auto Controller switched on, slave controller in “Cascade”
operating mode
manual Manual access to control output
Actual value TEMP-1 Nominal value of the process value in its physical unit,
e.g. °C for temperature, rpm for revolutions per min-
ute, pH for pH value, etc.
Target Setpoint Setpoint of process value in the physical unit, e.g. °C
for temperature
Controller Out Display of controller output in %
output
Alarm Alarm Enter alarm limits (high limit, low limit) and alarm
Parameter parameter status (enabled, disabled)
Profile Profile Param. Possibility to input a time-dependent setpoint profile
Parameter (max. 20 spikes)
Function key Access to controller parameters (with password) for
cascade controllers: Selection of slave controller
Function key ok Confirm entries with “OK”

17.4 Setpoint Profile Most control loops can be operated with time-dependent setpoint profiles (Control
Loop Profiles). The profile is input into a table using the operator terminal. Jumps and
ramps are possible in the profile; however, a profile can have a maximum of 20 spikes.
You can start and stop profiles at any time.
The elapsed time is displayed for started profiles.
Accessing Operator Screens

t Select the appropriate controller in the “Controller” menu.
t Call up the operator screen via the field “Profile Param.”.
Operator Screen

Add Adding a profile spike
off Setpoint profile not active
profile Setpoint profile has been started and is being processed
Setpoint [PV] Display of the current controller setpoint in the physical
units of the process value, e.g. °C for temperature
Elapsed Time h:m:s Display of the elapsed time since the profile start in
[hours:minutes:seconds]
Graphical display of the elapsed time on the profile
screen
No. 1-20 Number of the profile spike
Time h:m:s Input of the time for the profile spike
Setpoint [PV] Input of the setpoint for the profile spike in the physical
units of the process value, e.g. °C for temperature
Del Deleting a profile spike

17.4.1 Operation − We recommend that you create a sketch with spikes and associated setpoints for
your profile (see example). The time and setpoints to be programmed can be read
directly from the spikes entered into the sketch.
− A profile must contain at least one profile spike with a non-zero time in order to
be started.
17.4.2 Special Notes − When starting the setpoint profile, the controller mode will automatically be
switched to “profile” in the “Controller” menu.
− If you do not input the time “00:00 h:m” for the first spike, the system will use the
current setpoint as the starting time.
− In the case of a setpoint jump, the same time is programmable for both spikes.
− When starting a “DO (pO2)” profile, whichever profile for “STIRR”, “AIR”, or “PRESS”
has been started will be automatically stopped and switched to “cascade” mode.
17.5 General Controller For optimum adaptation of the controller to each control segment, the controller
Parameterization parameters can be changed using the parameterization screens:

MIN, MAX Value in % Minimum and maximum output limit for the
controller output
DEADB Value in °C Dead zone setting (PID controller only)
XP, TI, TD Value in %, sec PID parameters (PID controller only)

Parameterization screens are accessible after selecting in the controller
operator screen and password entry. DCU systems are configured at the factory with
parameters that ensure the stable operation of the control circuits in the bioreactor.
Factory-configured parameters can be found in the customer-specific configuration
documentation.
As a general rule, it is not required to change the control parameters. The exceptions
are controlled loop paths, the behavior of which is strongly influenced by the process,
e.g. pH and DO control loops.
17.5.1 Output Limits You can limit the controller output for the target value generator and PID controller
downwards (MIN) and upwards (MAX). In this way you can avoid unintentional,
extreme control element controls or limit the target value range for the slave
controller during cascade control.
− The limits are entered in the MIN (minimum limit) and MAX (maximum limit) fields.
The setting is made relative to the overall controller range in %.
− The following limits apply to the full modulation of the controller output:
− One-sided controller output: MIN = 0 %, MAX = 100 %
− Split-range controller output: MIN = -100 %, MAX = 100 %
17.5.2 Dead Zone A dead zone can be set up for PID controllers. If the control tolerance remains within
this dead zone, the controller output maintains a constant value and | or is set to
zero (pH controller). If the nominal values fluctuate stochastically, the dead zone
enables more stable control operations with minimized control element movements.
For controls with split-range outputs, this prevents oscillation of the controller
output (e.g. constantly changing acid | alkaline proportioning on the pH controller).
− The dead zone is displayed in the DEADB field or configured in the associated
submenu. Example for pH controller:
Set dead zone: ± 0.1 pH
− In that case, the control loop is inactivated at nominal values between 5.9 pH and
6.1 pH.

17.5.3 Controller Parameter-
ization Menu Screen
MIN % Minimum output limitation, limit value for switch to
upstream slave controller
MAX % Maximum output limitation, limit value for switch to
downstream slave controller
DEADB pH Dead zone in the unit of the process value
XP % P share (proportional range); signal amplification of
the control response proportional to the input signal
TI sec Integral portion; time function. With a higher I portion
control will react more slowly (and vice versa)
TD sec Differential portion: Damping, greater D portion,
damps the controller response (and vice versa)
OUT Controller output 1 (only in configurations in which
the switching of the output is provided)
OUT2 Controller output 2 (only in configurations in which
the switching of the output is provided)
17.5.4 PID Parameters The PID controllers can be optimized using the PID parameters “XP”, “TI” and “TD”.
The implemented digital controllers run according to the position control algorithm.
They allow structural toggles (P, PI, PD, PID) and changing the parameters during
ongoing operations.
− The controller structure can be configured by setting individual PID parameters to
zero:
P controller: Æ TI = 0, TD = 0
PI controller: Æ TD = 0
PD controller: Æ TI = 0
PID controller: All PID parameters defined
17.5.5 PID Controller Knowledge about control technology is prerequisite in order to be able to optimally
Optimization tune a PID controller to a controlled loop path; otherwise empirically tested tuning
methods (e.g. Ziegler Nichols) can be found in the pertinent literature. As a general
guideline:
− Only switch the D portion (TD) if the nominal values are relatively stable. For
stochastically variable actual values, the D portion makes fast, large changes to
the output. This causes unstable control.
− As a rule, the TI : TD ratio should be around 4 : 1.
− Periodic oscillations in the control circuit can be counteracted by increasing XP
and | or TI / TD.
− If the adjustments are too slow after target value jumps and | or in the case of
nominal value drift, you can lower Xp and | or TI | TD.

17.6 Temperature Controller The temperature control works like a cascaded regulation. The TEMP controller uses
the temperature measured in the culture vessel as a master value and acts on the
mode of the JTEMP slave controller. JTEMP's output controls the assigned control
elements through pulse-duration modulated or constant outputs in the split-range
operation.
Associated control elements can include:

− Electrical heaters in the temperature control circuit; valves on the steam supply of
steam-heated heat exchanger
− Valves of the cooling water supply(ies)
When the value approaches the setpoint, the guide controller switches the controller
structure from “PD” (starting condition) to “PID”, preventing overshoot. In the
temperature control circuits, like on bioreactors, a digital output also switches off the
circulating pump as well as the heating protection when the temperature controller is
switched off.
Master Controller TEMP operator screens
Refer to [¨ Section “17.3 General Controller Operation”] for notes on the fields,
entered values and entries.
Observe the maximum permissible temperatures of the component groups and

fixtures your bioreactor is equipped with.

The temperature cascade controller is operated from the master controller. You can
only change target values and operating modes on the master controller (TEMP).
All operations of the slave controller (JTEMP) are triggered automatically.
− For routine operation, you only need to configure the master controller (TEMP)
(set value, mode, and alarm limits).
− Direct configuration for heating and cooling can be done on the slave controller
(JTEMP) when the master controller TEMP is turned off (”manual” mode).
17.6.1 Special Notes − In “auto” mode of the TEMP master controller, the JTEMP slave controller
automatically switches to “cascade” mode. In the “off” setting of the master
controller, the slave controller is also automatically “off”.
17.7 Speed Regulator The DCU speed controller function works like a target value generator for an external
motor controller, which controls the speed of the stirrer motor. Operator entries,
output of the analog setpoint signal for the motor controller and the display of the
speed signal from the controller are all done on the DCU system.
When the stirrer speed controller function is switched off, an additional digital
output also modulates the drive protection. If the system has a DO (“pO2”) controller,
the speed controller function can be modulated as a slave controller in the DO
cascade control loop.
“Main” Menu operator screen

17.7.1 Special Notes
Depending on the vessel type, size, and equipment, only a certain maximum speed
is permitted. Higher speeds can damage vessel attachments. Vessels can become
unstable and move across the surface of the floor. Note the maximum permissible
speed for your bioreactor [¨ configuration documentation of the DCU system].
Culture vessel Maximum stirrer speeds BIOSTAT® C+

5L 1500 rpm
10 L 1500 rpm
15 L 1000 rpm
20 L 1000 rpm
30 L 600 rpm
If the MIN | MAX setting is changed after a system reset, you must reset the new
limits to the range permissible for the bioreactor.
When inputting MIN | MAX output limits or making direct entries into the OUT field,
the permissible speed controller range must be considered.
− Example: when selecting the speed control MIN | MAX 0 - 100% for speed range
0 - 2000 rpm and 1000 rpm as a permissible max. speed, a value of “OUT”:
MAX 50 % must be configured.

In addition to its function as a single controller, the stirrer speed controller can also
be used as a slave controller in DO (“pO2”) cascade control.
17.8 pH Controller The pH control normally works with PID control characteristics. It controls correction
medium pumps for acids and bases and | or proportioning valves or mass flow
controllers for CO2 in split-range mode using pulse-width modulated outputs. This
enables bilateral control.
− The negative controller output acts on the acid pump (or the CO2 feed) and the
positive output on the base pump.
− The pH controller does not activate the control signals until the control deviation
is located outside of a configurable dead zone. This prevents any unnecessary acid/
alkaline proportioning.
pH Controller Operator Screens
Refer to [¨ Section “17.3 General Controller Operation”] for notes on the displays,

17.8.1 Operating Notes A DEADB dead zone can be entered in the pH controller parameterization screen.
The controller remains inactive as long as the measured value remains within the
dead zone around the setpoint.
Set target value: ± 6.0 pH
In that case, the control loop is inactivated at nominal values between 5.95 pH and
6.05 pH.
17.8.2 CO2 Supply-driven For bioreactors for cell culture, a CO2 valve or a CO2 massflow controller can work as
pH Control the control element of the pH control in place of the acid pump.
17.8.3 Special Notes − The pH controller output “–Out” normally controls the acid pump with a negative
output signal (0 to –100%).
Correspondingly, the controller output “+Out” controls the base pump with a
positive output signal (0 - +100%) and adds base.
− In configurations for cell culture, the output “–Out” can be connected to the
CO2 feed.
After switching to “CO2”, the output controls the CO2 valve (or the massflow
controller of the CO2 segment) to introduce CO2 into the culture vessel.
− For special configurations, the acid or alkaline pump can be assigned to substrate
controllers if they are not needed for pH regulation.
To do this, “-Out” must be set to “None” (instead of “Acid” or “CO2”) and “+Out”
must also be set to “None”.
− When activating the modes “auto” or “manual”, the filling counters “ACID-T” /
“CO2-T” and “BASE” are automatically connected in mode “Totalize”.
17.9 DO Control Methods
17.9.1 DO Controller The DCU system features various methods of DO control. Which of them is possible,
required or sensible for the controlled terminal device depends on the configuration
or process.
− When aerating with air, either the oxygen portion can be reduced by adding
nitrogen, or the air can be enriched with oxygen.
− The overall gas flow can be controlled using a flow controller.
− The mixture can be influenced e.g. by controlling the agitator speed.
− Cell growth can be influenced by adding substrate.
The DO control works like a cascaded regulation. The output of the DO controller
(master controller) modulates the setpoint input of the slave controller, which then
acts on the control element (e.g. the valves or MFC for N2 or O2 or the agitator).
The following control strategies are possible:
− 1-stage control cascade, i.e. the DO control only affects one of the available
setting variables.
− Up to 4-stage control cascade, during which the DO control is affected up to 4
setting variables according to their priority.

A range (MIN | MAX) can be defined in the DO controller, in which the DO controller
defines the target value for each slave controller. In multi-stage cascaded regulation,
the output of the DO controller modulates the slave controller after sequential
switch-on as follows:
− The DO controller acts on the slave controller with the priority 1 (Cascade 1) and
defines its setpoint. The slave controller 2 receives the target value defined in the
DO controller as “MIN”.
− When the preset target value of the 1st slave controller reaches its maximum, the
output of the DO controller switches to the target value input of the 2nd slave
controller (Cascade 2) after an adjustable delay time “Hyst.” and defines the
following target values:
− Slave controller (cascade) 1: with defined maximum
− Slave controller (cascade) 2: controlled output of the DO controller
− This sequence continues for the other control elements according to the
predefined priority “Cascade #”.
− If the need for oxygen drops, the controllers are reset to in the reverse order.
In this way, the control can regulate the DO value in-process, even if there are
considerable fluctuations in the need for oxygen in the culture. In order to still be
able to additionally optimally adapt the control to the behavior of the controlled loop
path, the PID parameters of the slave controller are parameterizable independently of
one another.

DO (“pO2”) Cascade Controller Operator Screens
The operator screen also include the following input fields:

Field Value Function, display, entry required
Setpoint % sat Preset Target Value in the Master Controller
Setpoint Preset setpoint for the slave controller in the cascaded
Cascaded regulation, in the sequence of the priority predefined
Controller in the parameterization screen:
N2 N2 Supply Controller (Proportioning Valve)
AIR_SP Controller for Massflow Controller
O2 O2 Supply Controller (Proportioning Valve)
STIRR Speed regulator
SUBSA Corrective solution
OUT % Status of the slave controllers for cascade control with
actual value of controller output

DO (“pO2”) Cascade Controller Parameterization Screen

DEADB % Dead Zone Entry
Cascade # [Controllers] Slave controller with the relevant parameters
MIN % Minimum output limit, corresponding to the minimum
target value for the slave controller.
MAX % Maximum output limit, corresponding to the maxi-
mum target value for the slave controller.
XP % P share (proportional range); signal amplification of
the control response proportional to the input signal
TI sec Integral portion; time function. With a higher I portion
control will react more slowly (and vice versa)
TD sec Differential portion; damping, greater D portion,
damps the controller response (and vice versa)
Hyst. m:s Lag time for switching between the slave controllers.
Mode off / Slave controller operating mode after switching off
auto the DO controller.

17.9.1.1 Operation of the t Select the slave controller according to the desired priority in the “Cascade
Multi-stage Cascade Parameter DO” submenu.
Controller t Set each minimum and maximum controller target value limit for the selected
slave controller using the output limits MIN, MAX in the parameterization screen
of the DO controller (see previous illustration)
t When switching on the DO controller, the slave controller modulated by the DO
controller is displayed as “active”.
17.9.1.2 Special Notes − In modes “auto” and “profile” of the DO controller, the selected slave controllers
are automatically switched to “cascade” mode.
− In mode “off” of the DO controller, the selected slave controllers are also
automatically switched to “off”.
− Switching from slave controller 1 to the downstream controller and vice versa is
not done until the respective output limit for the time span defined in the “Hyst.”
field of the parameterization screen has been over- or undershot.
After this time has elapsed, check the switch conditions once again and only
switch back if they have been met.
− An inverted control direction for slave controllers, such as the substrate controller,
can be achieved by inverting the setpoint limit (MIN > MAX).
− The DO master controller always uses the MIN | MAX limits of the respective slave
controller as the working range.
− The difference between MIN and MAX must always be more than 2% of the
specific measurement range.
17.9.2 Advanced DO Controller The advanced DO controller monitors and regulates the DO in the bioreactor or in the
device controlled for which the DCU4 system was designed.
The controller acts as the master controller in the DO control cascade. It acts on a
configurable selection of slave controllers for the intake of media or to control
actuators that influence the DO in the process. Examples of such media include gases
like N2, air, O2 or nutrient solutions. The DO value measured in the process depends
on the media introduced, the oxygen consumed by cell growth and cell metabolism
and material distribution from mixing.
The master controller works as a PID controller with configurable control behavior.
It uses the DO measured at a measurement point (up to two measurement points can
be selected) as the actual value. In case of deviation from the setpoint, the master
controller sends an output signal to the slave controllers connected in cascade. Due
to the variety of possible slave controllers, the output signal is relative to the control
range 0 to 100%.
One configuration can include up to six slave controllers, of which five can be
selected simultaneously for the control cascade. They control their actuators using
analog or digital output signals. Each slave controller can be assigned up to five
setpoints in the physical units of the set value, dependent on the output “Out” of the
master controller. The controller operator screen shows this graphically as a polygonal
curve above the output “Out”.
In comparison with conventional DO control cascades, the advanced DO controller

supports the parallel operation of the slave controllers, that is, all actuators are
controlled simultaneously. In combination with the determination of multiple
setpoints dependent on the “Out” of the master controller, this results in an easy-to-
understand and convenient to operate DO cascade control.

DO (“pO2”) Controller Operator Screen

Settings for the Advanced DO Controller
Operating display and input window for the master controller

Mode Selecting the mode of the controller
[ off ] − Controller switched off, output on stand-by [¨ configuration]
[ auto ] − Controller active, controls the actuator if necessary
[ manual ] − Manual access to controller output
DO Display of the DO
Setpoint % Setpoint; in % relative to the control range 0 to 100 %
Out % Current controller output; in % relative to the control range 0 to 100 %
Access to the parameterization menu via standard password
[¨ see Section “20.8 Password System”]
[ Cascade Param. ] Access to the selection menu for the slave controllers, via standard password
Alarm PRESS Alarm monitoring settings
High limit % − Upper alarm limit
Low limit % − Lower alarm limit
Alarm state − Status: Alarm monitoring activated (enabled) or deactivated (disabled)
Operating Menus for Configuring Slave Controllers

N2-SP1 tag Slave controller assigned to this channel (order in the cascade)
N2, O2, AIR etc. tag − Media feed (gas, substrate) or function (e.g. speed controller)
SP etc. tag − Feed to the culture vessel or bag, e.g. sparger or overlay
1, 2 etc. # − The unit assigned to the controller output, e.g. culture vessel 1 to 6
End mode [ off ] Mode for slave controllers when the master controller is “off” or “disabled”; mode
[ auto ] restored after emergency shut off or power-on
Mode [ disable ] Manually switchable mode of the slave controllers (only available when the master
[ enable ] controller is in operating state “off” or “disabled')
Example: Input (modification) of the DO setpoint
Since the selection of slave controller can be changed according to process

requirements, the setpoint of the DO controller output is set in %) relative to the
control range. The slave controllers control their actuators with setpoints in their
physical units.
t Press DO (“pO2”) in the “Controller” menu.

t Press “Setpoint” and enter the password.
Access is password-protected in order to prevent unauthorized changes
[¨ see Section “20.8 Password System”].
t Use the numeric keypad to enter the set value.
Confirm with “OK”.

t Press the function key of the slave controller to be configured, e.g. “N2-SP1”.
Enter up to five set values that depend on the “Out” output of the master
controller. The settings are graphically displayed with a polygonal curve.
t Activate the controller by switching to the “auto” mode and confirming with “OK”.
17.9.3 Parameterizing the
Master Controller
Elements of the Parameterization Screens

Out % Current controller output “Out”, in % of maximum control
range
MIN % Minimum output, within 0 ... 100% of the control range
MAX % Maximum output, within 0 ... 100% of the control range
DEADB [PV] Dead zone; pressure regulation remains inactivate as long as
DO (“pO2”) deviates from the setpoint by less than DEADB
XP % P share (proportional range); signal amplification of the
control response proportional to the input signal; in % of
the measurement range
TI s Integral portion; time function of the control response.
With a higher I portion, the controller will react more slowly
(and vice versa)
TD s Differential portion; damping of the controller. With
a greater D portion, the controller response is reduced
(and vice versa)

Parameterization of the DO Master Controller
Normally you only change the parameters “MIN”, “MAX”, and “DEADB”:
t In the “Controller” menu, select DO (“pO2)” of the corresponding assembly to be
configured and open the controller operator screen.
t Press the parameter key and enter the password.
[¨ see Section “20.8 Password System”].
t Select the parameter to be set (”MIN”, “MAX”, or “DEADB”), enter the value and
confirm with “OK”.
Setting the “P”, “I”, or “D” Controller Parameters:
The adaptation of PID controllers requires knowledge of control theory. The setting
options listed here are rough guidelines.
Only qualified personnel should carry out controller optimization.
Depending on the process (e.g. stability of gas intake or actuator), it may be necessary
to change the parameters “P”, “I”, or “D” to adapt control behavior. You can test the
following changes:
− If the measured DO value (process value) oscillates around the setpoint and does
not stabilize, you can reduce the “P” portion.
If the actual value only approaches the setpoint very slowly or does not reach it,
you can increase the “P” portion.
− With a low “I” portion, the controller will react more quickly; as the “D” portion
falls, it will react more strongly to setpoint deviations. However, this can induce
the controller to tend to overshoot.
By increasing the “I” portion, we make the controller react more slowly, and by
increasing the “D” it will react more weakly to deviations in actual value.
This will make the controller response (the control behavior) more sluggish.

17.9.4 Selection and
Configuration of the
Slave Controllers

Elements of the Operator Screens for Selection and Configuration
Cascade # Slave controller to be assigned to the “Cascade #” position; up to 6 slave controllers are possible
[¨ configuration, specification]; up to 5 slave controllers can form a control cascade
N2, O2, AIR etc. tag Feed of media (gases, substrate) or actuators (e.g. drives)
SP, OV, FL etc. tag Introduction to control segment (e.g. sparger “SP”, headspace aeration “OV” in the culture vessel or
container, massflow controller “FL”)
1, 2 etc. # The unit assigned to the controller output, e.g. number 1 ... 6
Out % Output signal “Out” from master controller in control range 0 to 100% to which the setpoints of
the slave controllers should be assigned
Setpoint [PV] Setpoint of the slave controllers in their physical unit
End mode [ off ] Mode for slave controllers when the master controller is “off” or “disabled”; mode restored after
[ auto ] emergency shut off or power-on
Mode [ disable ] Manually switchable mode of the slave controllers (only available when the master controller is in
[ enable ] operating state “off” or “disabled')
Selection of Slave Controllers

t Activate “Cascade Param.” to open the submenu for selection of slave controllers
and change the previous selection.
t Enter the password.
[¨ “Appendix” in the DCU4 system manual].
t Press the key for position “Cascade #” for which another slave controller should
be selected or the existing one deselected.
Changes to a controller “Cascade #” deletes the subsequent selection. You must
reassign all downstream controllers.
Since the slave controllers control their actuators simultaneously, the order of the
controllers has no effect on the control.
Configuration of Slave Controllers

t Activate the function key of the slave controller you want to configure,
e.g. “AIR-SP1”.
t Enter the password.
Access is password-protected in order to prevent unauthorized changes [¨ see
Section “20.8 Password System”].
t In the “Setpoint” column, activate the key for the “Out” segment of the master
controller you want to assign a setpoint. Enter the setpoint that should
proportionally act in the control cascade, in the physical unit of the actuator.
t Enter the setpoint for the other “Out” segments. After closing the submenu with
“OK”, the setpoints are graphically displayed as a polygonal curve above the “Out”
of the master controller.
t Activate the submenu for the other slave controllers and enter their set values for
the “Out” segments of the master controller.

17.9.5 Special Notes The slave controllers work as long as the master controller is active, that is, in “auto”
or “manual” mode. After the master controller is turned off (in state “off”), the slave
controllers can be operated manually, either individually or together in the selected
combination.
The behavior of the master controller is based on sampled settings for the delay time
and switching hysteresis. These settings are determined internally and not accessible
for user modification. If necessary, they must be changed in the configuration.
The following settings are saved for the master controller and slaves:
− The setpoint
− The settings for alarm monitoring
− The PID parameters for the master controller and slave controllers
− Their settings relative to the output of the master controller
That means that these settings are available after a power outage or after the DCU4
system or the controlled device is turned off. They will be restored for the next
process after power returns or the controller is switched back on.
A reset of the DCU4 system [¨ see the chapter “19. “Settings” Menu”] restores the
factory settings. You must therefore store process- or user-specific settings before
the reset if you want to use them again later.
After loading a new system configuration, the DCU4 system initially starts up with
the factory settings. Here, too, you must reenter any process- or user-specific
settings.
17.9.6 Instructions for Use With an appropriate setup of setpoints for the slave controllers, they can work in a
conventional sequential control cascade. Example:
t Give “N2” a setpoint in the range “Out” = 0 to 20%, with the maximum at 0%.
t Give “AIR” a setpoint in the range “Out” = 0 to 20%, with the maximum at 20 %.
Leave “Out” constant for 20 to 100 %.
t Set “O2” between “Out” = 20 to 40%, with the maximum at 40%. Leave “Out”
constant for 40 to 100 %.
t Set “STIRR” between “Out” = 0 to 40% and increase to a maximum at 60 %.
Leave “Out” constant for 60 to 100 %.
t Leave “Substrate” constant in the range “Out” = 0 to 60% and increase to a
maximum at 80%.
y This activates the slave controller in the sequence shown, based on the deviation
between the actual and setpoints and the output signal of the master controller.
If the actual value approaches the setpoint, the slave controllers switch back in the
reverse order.

Examples of Applied Control Strategies
The examples are based on the control of mass flow controllers in the gas feeds.
Control strategies, e.g. O2 enrichment, exclusive flow, or gas flow ratio, can be
implemented by the selection and configuration of control cascades:
O2 Enrichment
t Select “AIR” and “O2” as slave controllers.
t For “AIR”, set up a constant setpoint over the entire control range “Out” = 0 to
100%.
t For “O2”, set up the lower (minimum) setpoint up to “Out” = 40% and the upper
(maximum) setpoint starting at “Out” = 60%.
y This yields enrichment with oxygen starting at “Out” = 40%.
Setting
Aeration Rate (%)
DO controller output (%)
touch panel
DO Out (%)

Exclusive Flow
t Select “N2”, “AIR” and “O2” as slave controllers.
t For “N2”, set the maximum setpoint at “Out” = 0% and the minimum at “Out” =
20%.
t For “AIR”, set the minimum setpoint at “Out” = 20%, the maximum at “Out” = 40%,
and all other “Out” up to 100%.
t For “O2”, set the minimum setpoint at “Out” = 40%, the maximum at “Out” = 60%
and all other “Out”.
y This configuration dispenses N2 at a controller “Out” below 20%. Air is added for a
controller “Out” of 20% or more, and the oxygen contribution rises starting at
“Out” = 40% by the introduction of O2.
Setting
Aeration rate (%)
touch panel
DO Out (%)
Setting
Aeration rate (%)
touch panel
DO Out (%)
Setting
Aeration rate (%)
touch panel
DO Out (%)

Gasflow Ratio Air | O2 (Total)
The “Gasflow Ratio (Total)” aeration strategy is only possible with “AIR” and “O2”
as slave controllers and if the gas feeds have massflow controllers as actuators
[¨ configuration, PI diagram].

t Set the minimum “AIRFL” setpoint for “Out” = 0 to 40% and a setpoint (not the
maximum) starting at “Out” = 60%. This gives the DO that should be achieved by
adding air.
t Set the minimum “O2” setpoint for “Out” = 0 to 40% and increase the setpoint by
a certain amount starting at “Out” = 60%. The increase yields the DO content that
proportionally should be achieved by the addition of oxygen.
y The added air is enriched with oxygen in the range “Out” = 40 to 60% of the DO
setpoint, with a maximum oxygen feed in the range “Out” = 60 to 100 % of the
DO. Air and oxygen portions add up to a relative maximum “Total” = 100%.
Setting
Aeration rate (%)
touch panel
Setting
Aeration rate (%)
touch panel

Gasflow Ratio Air / O2 (Ratio)
The “Gasflow Ratio (Ratio)” aeration strategy is only possible with “AIR” and “O2”
as slave controllers and if the gas feeds have massflow controllers as actuators
[¨ configuration, PI diagram].

t For “AIR”, set up the maximum set value up to DO (“pO2”) of “Out” = 40% and the
minimum set value starting at “Out” = 60%.
t For “O2”, set up the minimum set value up to DO of “Out” = 40% and the
maximum set value starting at “Out” = 60%.
y This means that in the DO setpoint range “Out” = 0 to 40%, only air is added, that
is, only the air feed regulates the DO. In the range “Out” = 40 to 60%, the portion
of air reduces to a minimum and the oxygen portion increases to its maximum. In
the range “Out” = 60 to 100%, only the oxygen feed regulates DO.
Setting
Aeration rate (%)
touch panel
DO Out (%)
Setting
Aeration rate (%)
touch panel
DO Out (%)

17.10 Gas Filler Controller Gas filling controllers control valves for the corresponding gas feeds, e.g. “AirOV-#',
“AirSp-#', “O2Sp-#', “N2Sp-#', “CO2OV-#”, or “CO2Sp-#” and introduce gases into
the aeration segments “Overlay” or “Sparger”.
The controllers normally work as slave controllers for the DO (“pO2”) or pH control
loop. When the DO control is switched off, they can be used as target value
controllers:
Depending on how the system is configured, gas filling controllers are available as
slave controllers and|or setpoint generators.
Operating Menus
17.10.1 Operating Notes To operate the gas filling controller as a setpoint generator, the master controller
must be switched off. Check its operating mode in the “Main” or “Controller” menu,
and switch the mode of the master controller to “off” if it is active.
t Select the “Main” or “Controller” view in the detail view “1”... in which you want to
set up the gas filling controller.
t Select the function key with the current display of the setpoint “0.0 lpm”. Enter
the setpoint in the window with the numerical keypad.
t Set the alarm limits, if needed, and activate alarm monitoring.
t Select the function key for the operating mode and select the “auto” operating
mode.
t Press “OK” to activate the controller.

17.10.2 Special Notes t Select the setpoint of 100% to configure the flow rate on the flow meter and
calibrate the filling counter (if the calibration function is included in the
configuration). Oxygen then flows continuously in the air supply.
t For manual gas supply, select the desired setpoint in the range 0 to 100 %.
y When activating the “auto” mode on the master controller, the gas filling
controller automatically switches to “cascade” mode. In that event, settings in the
gas filling controller are not possible or will be ignored.
17.10.3 Gas Flow Controller Note the specifications for the measurement | control range of the aeration rates of
the bioreactor. When a bioreactor is operated with overpressure, the counter pressure
might cause the maximum aeration rate not to be reached.
Gas flow controllers control the massflow controllers of the assigned gas segment
(”GAS-SP” or “GAS-OV”) [¨ PI diagram]. The massflow controller makes it possible to
aerate the reactor vessel with a continually changing gas flow.
The gas flow controller normally operates as a slave controller in the DO (“pO2”)
cascade control circuit. The master controller (DO controller) controls the massflow
controller in accordance with the sequence in the control cascade, using a continuous
output signal.
The gas flow controller can be deselected in the master controller. It is then available
as a setpoint generator. It controls the massflow controller using an analog setpoint
signal.
Operating and Parameterization Menu

Gas Flow Controller Settings
Operator screen
Function key Mode Input of the controller mode
Manual − Manual access to control output
Auto − Automatic operation, control with a predefined
target value
off − Controller switched off, output on stand-by
[¨ configuration]
XYZ_FL ccm | lpm Current total gas flow
Setpoint ccm | lpm Target value for the flow controller
Access to the parameterization menu with a password
Out % Current controller output

Alarm GASFL Alarm monitoring settings
HiLim % − Upper alarm limit
LoLim % − Lower alarm limit
Alarm state − Status: Alarm monitoring activated (enabled) or
deactivated (disabled)
Parameterization screen
MIN % Lower output limit, setting range 0 to 100% of the
control range
MAX % Upper output limit, setting range 0 to 100% of the
control range
Out The controller output is assigned to the control
element (if implemented)
Special Notes
Follow the instructions regarding the “Parameter Settings in the System” in the
“Configuration Documentation”.
− MIN | MAX output limits are entered in % of the control range of the gas feed.
When entering values directly in the OUT field, take the measurement range for
the aeration rate into consideration.
− If the gas flow controller is a slave controlled in the DO control cascade, enter the
MIN | MAX values in the “DO controller” parameterization menu. The settings will
then act as a switching criterion for cascade control.
− Switching off the GASFL flow controller (select “off” and after an emergency shut
off due to overpressure) closes the control valve in the mass flow controller.

17.11 Foam and Level The input signal to the controller is a limit value signal generated by the measure-
Controllers ment amplifier to which the foam and | or level sensor is connected. This is activated
as long as the sensor is in foam or medium. The response sensitivity of the measure-
ment amplifier can be set on the operator screen of the controller.
The output of the controller modulates a correction medium pump and switches it on
and off periodically when a sensor signal is emitted. You can enter the pump run and
cycle time for repeated switching on and off on the controller operator screen.
This segment shows an example foam controller. Specifications in menus and settings
apply accordingly for the level controller.
Level Controller Operator Screens

Function key off Controller switched off
Auto Controller switched on
Manual Manual access to control output
Cycle h:m:s On and off time of the control element output
Cycle time in [minutes : seconds]
Pulse h:m:s On time of the control element output
Filling time in [hours:minutes:seconds]
Sensitivity Low…High Trigger sensitivity for the sensor
Function key Access to the parameterization menu
(with a password)
Alarm Alarms Enter alarm limits (high limit, low limit) and
Parameter Param. alarm status (enabled, disabled)
High Limit % Upper alarm limit
Low Limit % Lower alarm limit

17.11.1 Displays
Probe signal off Signal on, output auto – off
Output manual - on Signal on, output auto - on
17.11.2 Operation t Set the cycle time “Cycle” and the dispensing time “Pulse” according to process
requirements.
t Select the trigger sensitivity “Sensitivity” of the sensor:
“Low”, “Medium Low”, “Medium High”, or “High”.
To prevent proportioning errors resulting from leakage currents and sensor
growth, you should set the response sensitivity as low as possible.
t Switch the mode to “auto”.
In “manual” mode, the pump for long-term operation can be switched on or off.
17.11.3 Special Notes − The measurement amplifier is equipped with a response lag time mechanism
(approx. 5 sec), that prevents activation after splashing liquid.
− Switching to the “auto” or “manual” mode automatically also activates the filling
counter “AFOAMT” and|or “LEVELT”.

17.12 Gravimetric Filling The controller “FEED #F” is a precise gravimetric filling controller.
Control It is used a weighing system and an analog filling pump.
Because the control algorithm in the DCU system works directly with the weight
measured on the scale | balance, the gravimetric filling controller allows accurate
proportioning over days and weeks.
Operator and Parameterization Screens
17.12.1 Operation Operation with supply container and filling controller:

t Tare the scale to zero and set the vessel on the scale or hang the vessel or bag into
the weighing system.
t Enter the setpoint for the filling controller into the DCU system.
t Switch the mode of the filling controller to “auto”.
A negative weight readout on the scale | balance or on the DCU indicates the feed
volume.
17.12.2 Special Notes − The feed volume of the filling pump has an important influence on the controlled
loop path. That is why the pump throughput must be adapted to the required flow.
− For accurate proportioning, the working range of the controller output (”Out”)
must lie in the range from 15 to 90%. For that purpose, you can adapt the feed
range of the pump to the working range of the controller. You can use hoses with
a different diameter that offer the desired conveying range.

17.13 Filling Pump Controller To introduce nutrient solution, the filling pump controller can control an internal or
external pump. The controller function works as a setpoint generator, handles remote
control, and emits an analog setpoint signal for the pump.
Operator Screens
17.13.1 Special Notes − Matching connecting cables are available for certain pumps, like WM 101,
WM 323. Ordering information is available on request.
− Pumps from other manufacturers can be connected if they have an external
setpoint input 0 ... 10 V, 0 | 4 ... 20 mA.

17.14 Pump Assignment All controllers that can control pumps are assigned a pump. If the configuration so
specifies, the controller outputs can be connected to other pumps. However, only one
controller at the time can be connected to the corresponding pump.
If there are no external substrate pumps available, you can switch the substrate
controller to one of the internal pumps not in use.
Operator Screens

OUT Pump that works on the controller:
SUBSB External pump (signal to output “Substrate”)
ACID ACID pump (if released in the pH controller)
BASE BASE pump (if released in the pH controller)
AFOAM AFOAM pump (if released in the foam controller)
LEVEL LEVEL pump (if released in the level controller)
FO/LE FO/LE pump (if released in the FO/LE controller)
None No pump assigned, the OUT of another controller can
be assigned a previously allocated pump.
17.14.1 Operation To switch the assignment of a controller output to a pump, proceed as follows:
t Release the pump not in use by the other controller at its output “OUT”.
Example:
− Set the output “OUT” in the pH controller to “None”.
t In the substrate controller, assign the now-free pump under “OUT”.

Example:
− Set the output “OUT” in the SUBSB controller to “ACID_##”.
17.14.2 Special Notes The configuration of the DCU4 system must permit the desired assignment and switch
the pumps to the controller outputs. If not,
− Either no “OUT” switch is visible or selectable
− or the pump is hidden and cannot be selected, e.g. “ACID_##”.
If the pump switch is dimmed and cannot be selected although the configuration
allows switching, the assignment was not cancelled in the previous controller.

18. “Phases” Menu
18. “Phases” Menu
18.1 General Information Reactor-specific configurable phases can be implemented in the DCU system. This
type of program can, for example, control vessel sterilization, medium sterilization,
the sterilization of transfer systems, or the intermediate sterilization of the exhaust
filter. The parameters used in the procedure, such as temperature and time, can be
adjusted to specific requirements.
“Phases” Main Screen Operator Screens
Available Phases
Key, symbol Meaning, use
Sterilization Phases
Empty sterilization of the culture vessel:
− Intake and exhaust air are sterilized from the vessel
Full sterilization of the culture vessel:

− Intake and exhaust air are sterilized from the vessel
Pressure Hold Test of Culture Vessel
Phase Parameter Selection Key

By pressing the “Phase parameter” selection key in the corresponding phase, any
additional parameters needed for control of the process can be configured.
Access is password-protected in order to prevent unauthorized changes [¨ Section
“20.8 Password System”].
200 “Phases” Menu

18.2 Phase Process Control There are two different types of phase process control:
− Automatic process control
− Single-step control
Automatic Process Control

Automatic process controls, for example, are used for the sterilization of the vessel
and the connected peripheral assemblies. The entire sterilization procedure takes
place as a sequential control depending on time and events. The individual steps are
automatically followed in sequence.
− The necessary parameters “Sterilization temperature”, “Sterilization time”, and
“Fermentation temperature” are entered into the operator screen.
− You can start the sterilization from the operator terminal (and also stop it if
needed). The operator screen shows the active step and the elapsed sterilization
time.
− If the automatic procedure requires additional manual actions on the bioreactor,
e.g. the opening or closing of manual valves, the phase control displays a
corresponding messages when that step is reached. The program only continues
the phase when the manual action has been carried out and the message
acknowledged with the “OK” button.
Single-step Control
Single-step control is used for the intermediate sterilization of peripheral
attachments such as the exhaust air filter, for example. The sterilization program
specifies a practical order for the steps and the user confirms the steps with the “OK”
button as needed.
− In a single-step control, certain steps can also automatically expire using a timer
(e.g. the sterilization time you enter on the corresponding operator screen).
− You can also start single-step control from the operator terminal and confirm the
input with “OK”. If necessary, the procedure can also be stopped with “State: stop”.
The operator screen shows the currently activated step and, if necessary, the
sterilization time that has already elapsed.
“Phases” Menu 201

18.2.1 Status Displays During Both during automatic process control and in single-step control, the header line on
Step Control the operator screen shows the process status for the running program, e.g. “State:
Running”.
General Status Displays and Functions

State Input to start or stop the phase
Start − Starts the phase
stop − Stops the phase
step − Manual transition to the next step
Display the status in the program
Running − Program is running
Locked − Phase start not possible; another phase or another
recipe is active
Idle − Program not active
STEMP degC Sterilization temperature
FTEMP degC Process temperature
SJTEMP degC Sterilization temperature of double wall
STIME h:m:s Sterilization time in [hours : minutes : seconds]
Elapsed h:m:s Sterilization time elapsed in [hours : minutes : seconds]
MAXTIME h:m:s Max. sterilization time in [hours : minutes : seconds]
after the sterilization temperature is reached
Setpoint Table [PV] Input of process parameters

18.2.2 General Procedure t In the “Phases” menu after pressing the appropriate icon, select the phase desired
for Phase Control [¨ see Section “18.1 General Information”].
t Start the phase by pressing the “State” key and selecting the “start” mode.
t If manual actions are required, when the system requests it carry out the measure
and acknowledge the message by pressing the “OK” button.
y After ending the sterilization program, the operator terminal shows the alarm
message “Sterilization finished” in the last step.
t Acknowledge the alarm message with the “Acknowledge” button to deactivate

the alarm.
y You can stop the automatic phase procedure at any time with “State: stop”.

18.2.3 Displaying
Check the messages in the “Condition” field carefully. Only acknowledge the message
Conditions
with the “OK” button after all the requirements have been met.
In the “Condition” field, the conditions for the current process step are displayed.
Some conditions displayed must be confirmed with the “OK” button after a thorough
test.
The exact description of the operating steps required can be found in the operating
manual for the system in [¨ Chapter “6. Operation”].

18.2.4 Special Notes
Faults in the operating sequence are displayed as alarms on the operator terminal.
Check the messages and correct the cause of the alarm.
Failure to observe the alarm message could result in damage to the device.
− When a phase is running, such as vessel sterilization, the operator terminal shows
the process status as “State: Running”.
− If no process time has been set, the process time automatically starts when the
sterilization program starts.
− Running phases can be stopped at any time. When vessel sterilization is stopped,
a cooling process is automatically started that cools the bioreactor to the preset
operating temperature for the process as quickly as possible.
− If necessary, a stopped sterilization program can also be restarted before the
operating temperature is reached.
− If the status display shows “State: Locked”, the phase is locked because another
phase, recipe, or process is active. The release for start only takes place when the
program that is active has ended.
18.3 Sterilization Phases

Danger of injury in the vicinity of the bioreactor!
− The vessel and in-situ sterilized components and lines are heated to the
sterilization temperature and are under pressure.
− Parts that are incorrectly installed or manipulated, as well as steam and | or hot
culture medium, can be explosively ejected.
− Scratches or hairline cracks on glass containers (correction medium and sampling
bottles) can impair their pressure resistance to the point that operational safety
can no longer be ensured for sterilization. Handle the culture vessel very carefully.
− Before starting the bioreactor, be sure that no personnel is within the danger zone.

− Use safety gloves when manipulating the valves.
− If sterilization is interrupted, wait until the bioreactor has achieved a safe state
(cooled to ambient temperature, pressure released) and then continue with work
on the vessel.
− While operating the bioreactor, be sure that no personnel is within the danger
zone.
The culture vessel sterilization is carried out in several steps in a defined sequence.
The operator terminal is used to set individual parameters (e.g. the sterilization
temperature), control the process of sterilization if needed, and read off the current
process status. Phases for full sterilization and | or empty sterilization can be
implemented for the sterilization process.

Operator Screen
Display of Steps and Conditions

Field Step Condition
actual Step Display of the sterilization step
---- − Sterilization program not active
MANOP − Preparation for sterilization
HEAT1 − Heat to 98 °C
HEAT2 − Heat to sterilization temperature
STERI − Sterilization procedure running
PV UNDER-LIMIT − Sterilization will stop until the process value
in question is within the limits
COOL1 − Cooling from the sterilization temperature
to 98 or 80 °C (depending on the bioreactor)
COOL2 − Cooling from 98 or 80 °C to the process
temperature (depending on the bioreactor)
READY − Process temperature reached
(message “Sterilization finished”)
END − Sterilization ended
next Step Display of the next sterilization step

Operation
To carry out culture vessel sterilization, proceed as follows:
t Prepare the bioreactor for sterilization as described in the operating manual.
t Carry out a pressure test or pressure hold test for the culture vessel
t If necessary, change the sterilization parameters (e.g. the sterilization temperature,
sterilization time, or process temperature to which the reactor will be cooled after
sterilization is complete).
y The predefined sterilization temperature is 121 °C. Increase this value only if the
attachment parts in the vessel such as sensors are suitable for higher temperatures.
y The preset sterilization time is 30 min. (Time the temperature is held at 121 °C).
If you do not achieve reliable sterilization within this time, you will have to
determined the time required on an empirical basis.
t Start the sterilization program by selecting the status “State: start”.
If manual actions are required, when the system requests it carry out the measure
and acknowledge the message.
t After the automatic sterilization is complete, acknowledge the message
“Sterilization finished” by pressing the “Acknowledge” button.
You can stop the automatic sterilization procedure at any time with “State: stop”.

18.4 Additional Phases
18.4.1 Pressure Hold Test of Culture Vessel The pressure test and | or pressure hold test should be carried out before each
sterilization of the culture vessel. The test ensures that all screw connections and
ports on the culture vessel are sealed.
Danger of injury in the vicinity of the bioreactor!

− The vessel and lines are pressurized. Attachments that are incorrectly installed or
being manipulated may be ejected.
− Before starting the bioreactor, be sure that no personnel is within the danger zone.
Display of Steps and Conditions

Field Step Condition
actual Step Display of the process step
---- Program not active
MANOP Manual operation required, prepare system for
the phase
PRESS Pressurization of the culture vessel including
assemblies
HOLD Pressure hold time
RLEAS System pressure relief
READY Process value reached (message “PHOLD finished”)
END End of process
next Step Display of the next process step
Test Time hh:mm:ss Pressure hold time in [hours : minutes : seconds]
Test pressure mbar Pressure for test [mbar];
access only through phase parameters
Release press mbar Pressure after pressure hold test [mbar];
access only through phase parameters
Elapsed hh:mm:ss Sterilization time elapsed in [hours : minutes :
seconds]
Operation
To carry out the pressure hold test, proceed as follows:
Prepare the system for the pressure hold test
t Activate temperature control and wait until the set target value has been reached.
t Start the pressure hold test.
If manual actions are required, when the system requests it carry out the measure
and acknowledge the message.
t After the pressure hold test is complete, acknowledge the alarm message by
pressing the “Acknowledge” button.
You can stop the automatic procedure at any time with “State: stop”.

19. “Settings” Menu
19. “Settings” Menu
Unauthorized settings can lead to injury!
The “Settings” menu permits changes to the system configuration. Malfunctions that
have unforeseeable impacts on safe operation can result from settings that are not
permissible or are unsuited for a certain terminal device.
Settings that impact safe operation are password-protected. Only trained and
experienced persons may change these settings.
− The standard password [¨ see Section “20.8 Password System”] may only be
disclosed to authorized users,
− the service password [¨ separate notice] only to authorized service technicians
and administrators
19.1 General Information In the “Settings” menu, the DCU system provides various functions for system
maintenance and troubleshooting:
− General settings like date, time, fail time, password -protected screen saver,
parameter settings for communicating with external devices (”Internet
Configuration”).
− Defining Process Values (PV) and their ranges and limits.
− Manual operation of digital and analog inputs and outputs or simulation
controllers, for example.
− Service function, e.g., for resetting the system (Reset) or to select the system
configuration on multiple configurations.
“Settings” Operator Screen
“Settings” Menu 209

Functions Available for Selection
Touch key Function
System parameters Change general system settings
[¨ Section “19.2 System Settings”]
PV ranges Configure measurement ranges for process values
[¨ Section “19.3 Measuring Range Settings”]
Manual Operation Switch process inputs and outputs to manual operation
[¨ Section “19.4 Manual Operation”]
External View status of externally connected devices, e.g. scales
[¨ Section “19.5 Externally Connected Devices”]
Service Service and diagnostic interventions
[¨ Section “19.6 Service and Diagnosis”]
System Information Displayed

Hardware Microbox Version of the DCU hardware
Firmware X.YY Version of the system's firmware
Configuration XX_YY_ZZ Version of the configuration
For inquiries about the system or for contacting the service department in the
event of a malfunction, please always state the firmware indicated here and the
configuration of your system.
19.2 System Settings Using the “System Parameters” touch key, you can change general system settings,
for example setting the real-time clock on the DCU system.
To open the “System parameters” submenu, you will need to enter the standard
password [¨ Section “20.8 Password System”].
210 “Settings” Menu

Time hh:mm:ss Displays the current time,
format: hour:minutes:seconds
Date dd.mm.yyyy Input current date, format: day.month.year
Beeper enabled | Turns acoustic signals on | off, e.g. alarm tones
disabled
Fail time hh:mm:ss Enter power outage time to tell system how to behave
when switched back on, format: hour:minutes:seconds
Power outage time < FAILTIME: The system continues to
run on the previous settings
Power outage time > FAILTIME: System switches
to default mode [¨ Chapter “12. System Behavior
During Start”]
Screen hh:mm:ss Enter the time of inactivity after which the screen saver
saver will be turned on, format: hour:minute:second
(00:00:00 = switched off)
Internet 16-digit bi- The DCU system's address in the IP network
Config nary number
Changes to “Date” and “Time” will only take effect in the first 5 minutes after the
DCU4 system is turned on.
19.3 Measuring Range Settings The beginning and end of the measuring range (”PV Ranges”) for all process values
can be changed in the “Settings” menu. Measuring ranges configured specifically to
devices or customer specifications are factory-set in the bioreactor
[¨ Configuration documentation].
Only personnel authorized to do so may change the menu settings.

To make settings in the menu, a “system password” needs to be entered
[¨ Section “20.8 Password System”].

Operator Screens
After pressing the “PV ranges” touch key and entry of the standard password,
the “Process Value Ranges” submenu opens:
By pressing the “Ch.” (Channel) touch key, the process values (ranges) can be
configured:

Ch. Channel
Process value 0 ... 100% % or physical unit
Min Minimum value
Max Maximum value
Decimal Point Decimal point display
Alarm Low °C Lower alarm limit in the physical unit
Alarm High °C Upper alarm limit in the physical units
Alarm disabled Alarm monitoring deactivated
enabled Alarm monitoring alarms active
Delay s Alarm lag time

19.4 Manual Operation When starting up operations and troubleshooting, all analog and digital process
inputs and outputs as well as DCU internal inputs and outputs can be switched to
manual operation (”Manual Operation” touch key).
− To open the “Manual Operation” submenu, you will need to enter the standard
password [¨ Section “20.8 Password System”].
− You can disconnect inputs from the external signal generators and preset input
values to simulate measuring signals.
− You can separate outputs from internal DCU functions and directly influence
them in the operator screen, for example to test the effect of certain settings.
Settings during manual operation have the highest priority; their effects on the
inputs and outputs of the DCU system supersede those of other functions.
Color Displays of Inputs | Outputs

If an input or output is in “Manual” operation, the display in the “Value” column has
a green background.
− If a controller is in cascade control mode, the display in the “Setpt” column has
a light green background (controllers only).
− If a phase is acting on an output, the display in the “Value” column has a turquoise
background.
− If an input or output is in “Auto” operation, the display in the “Value” column has
a yellow background.
− If an input | output is locked, the display in the “Value” column has a violet
background.
− If an emergency off (shutdown) is triggered during process, the displays of all
outputs in the “Value” column have a red background.
− If no function is accessing an input | output, the display in the “Value” column has
a gray background.
− If the process control system is accessing an output, the display in the “Value”
column has a white background.

19.4.1 Manual Operation t For manual operation, disconnect the digital input from the external signal
of Digital Inputs generator, e.g. limit value generator, and simulate the input signal by entering
“ON” or “OFF”.
Digital Inputs Operator Screen

Day Description Display of digital input
Port Description Hardware address
Value PV Display of signal level of switching state
0 V = off
5 V / 24 V = on,
Input for “AUTO” or “MANUAL ON | OFF”
operating modes:
“AUTO”: Normal operation, external input acts on DCU
“MANUAL”: Manual operation, manual specification of
digital input
A Display active status
I: on = turned on (signal level 24 V)
N: on = turned on (signal level 0 V)
off : turned off

AL Alarm state
A = activated
– = not activated
PV Switch status of the digital input
off = turned off
on = turned on
19.4.1.1 Special Notes The following signal levels apply to the switch status (status):
off 0V
on 5 V for DCU-internal inputs (DIM);
24 V for process inputs (DIP)
After working on the manual level, you have to switch all inputs back to the “AUTO”
operating mode. Otherwise, the function of the DCU system will be limited.
19.4.2 Manual Operation During manual operation, disconnect the digital output from the internal DCU
of Digital Outputs function and manipulate it directly.
For static digital outputs, e.g. controlling valves, switch the output on or off.
For pulse-width modulated outputs, set the switch-on ratio in [%] by hand.
Multiple functions may act internally on a digital output. After the field is selected,
the currently active function will be displayed in the VALUE column in the
corresponding submenu. If several functions are activated (e.g. on controller
outputs that interact with sterilization), the following priority applies:
Highest priority Shutdown
Manual Operation (Manual Level)
Locking
Sterilization (only reactors capable of in situ sterilization)
Pump calibration
Controllers, timers, sensors, scales | balances
Lowest priority Controllers, etc.

Digital Outputs Operator Screen

Day Description Display of digital input
Val off Switch status of the digital output
on off = turned off
nn % on = turned on
% = power-on ratio (0 ... 100 %)
for pulse-width modulation of digital outputs
operating modes:
“AUTO”: Normal operation, external output acts on DCU
digital output
A Display active status
I = turned on (signal level 24 V)
N = turned on (signal level 0 V)
off = turned off
Ty Upstream function
cl = controller
expr = logical function
– = without

SRC nn% | off Upstream controller output
Display of output value:
– off
– –100% … +100%
19.4.2.1 Special Notes The following signal levels apply to the switch status (status):
off 0V
on 24 V for process outputs (DOP, DO)
On pulse-width modulated digital outputs, the relative power-on time is displayed

and | or preset. The cycle time is defined in the specific configuration.
Example
Cycle time 10 sec, PWM* output 40%:
− Digital output 4 sec on and 6 sec off.
* PWM: Pulse-width modulation

19.4.3 Manual Operation You can disconnect all analog inputs from the external circuitry during manual
of Analog Inputs operation, e.g., a measurement amplifier and simulate them by entering a relative
signal level (0...100%).
Analog Inputs Operator Screen

Day Description Display of analog input
Value PV Input signal 0 ... 10 V or 0/4 ... 20 mA
Input for “AUTO” operation or “MANUAL ON | OFF”
PV Process value
Unit Physical variable

19.4.3.1 Special Notes For internal analog inputs (AIM), the physical signal level is always 0 ... 10 V
(0 ... 100 %).
For external analog inputs (AIP), the signal level can be configured between
− 0 ... 10 V (0 ... 100%)
− 0 ... 20 mA (0 ... 100%)
− 4 ... 20 mA (0 ... 100%)
During manual operation, only the relative signal level (0 ... 100%) of the analog
inputs is displayed or entered. The allocation to the physical value is a product of the
measuring range of the affected process value.

19.4.4 Manual Operation You can disconnect analog outputs from the internal DCU functions and influence
of Analog Outputs them directly using signals with a relative level (0 ... 100%).
Output signals have the following priorities:

Highest priority Shutdown
Manual Operation (Manual Level)
Locking
Lowest priority Controllers, etc.
Analog Outputs Operator Screen

Day Description Display of analog output, e.g. STIRR
Port Description Hardware address, e.g. 1A005
Value PV Output signal 0 ... 10 V or 0/4 ... 20 mA
operating modes:
analog output

Ty Upstream function
cl = controller
expr = logical function
− = without
PV Process value
19.4.4.1 Special Notes The physical signal level of the analog outputs (AO) can be configured between:
− 0 ... 10 V (0 ... 100%)
− 0 ... 20 mA (0 ... 100%)
− 4 ... 20 mA (0 ... 100%)
19.4.5 Manual Operation for You can simulate controllers in manual operation by entering a setpoint.
Controllers (”Control
Loops”)
“Control Loops” Controller Operator Screen

Day Description Display of controller, e.g. TEMP
PV Process value
Setpt Display of set value
Input for “OFF” or “AUTO” operating modes:
“OFF”: Controller is turned off
“AUTO”: Normal operation, setpoint for controller can be
configured
Setpt Display of set value
C Display of active cascade
0 = no cascade
1 ... n = specific cascade for cascade controller
Out Calculated output value
19.4.5.1 Special Notes After working on the manual level, you have to switch all inputs back to the “AUTO”

19.4.6 Manual Operation for You can disconnect counters from the external circuit in manual operation and
Counters (”Digital simulate them by entering a frequency.
Counters”)
“Digital Counters” Operator Screen

Day Description Display of counter, e.g. TEMP-1
Port Description Hardware address, e.g. 1DC1
Freq Display process value | configured frequency
Input for “AUTO” or “MANUAL” operating modes:
“MANUAL”: Manual operation, frequency is configured
PV Display of measured process value
19.4.6.1 Special Notes After working on the manual level, you have to switch all inputs back to the “AUTO”

19.4.7 Manual Operation of You can simulate sequences in manual operation (e.g. during startup or in case of
Sequence Control problems in the sequence execution during sterilization) by starting a sequence.
(”Phases”)
Sequence Control Operator Screen

Day Description Display of sequence, e.g. FVESS-1
State Display of sequence status | step
Start | stop of a sequence (”START” | “STOP”)
Continuation to next sequence step (”STEP”)
Step Display of current sequence step
19.4.7.1 Special Notes Type and number of sequence steps of individual sequences depends on the

19.5 Externally Connected The “External” menu button can be used to view and set the status of externally
Devices connected units (e.g. scales).
Only personnel authorized to do so may change the menu settings. To make settings
in the menu, the standard password needs to be entered [¨ Section “20.8 Password
System”].
After pressing the “External” touch key and entry of the standard password, the
“External System” submenu opens:
“External System” Operator Screen

Day Description Display of device, e.g. FEEDW-A1
Interface Description Display of interface
Alarm Display and configuration of alarm status:
enabled = activate alarm
disabled = deactivate alarm
Status Display of status of connected device (offline | online)

19.6 Service and Diagnosis Type and number of sequence steps of individual sequences depends on the
This operating level is only accessible for interventions by authorized service

technicians or associates of Sartorius Stedim.

20. Appendix
20. Appendix
20.1 Alarms The DCU system makes a distinction between alarms and messages. Alarms have
higher priority and are displayed first ahead of the messages.
20.1.1 Alarm Triggering When alarms are triggered, they automatically are displayed in a window that
superimposes all other windows. The color of the soft button alarm bell turns red.
The color of the alarm bell stays red as long as at least one unconfirmed alarm
remains in memory.
“New ALERT” Operator Screen
Closing the window:

− After pressing , the alarm is stored as an unacknowledged alarm “UNACK” in
the alarms list and the alarm symbol remains activated.
− The alarm window closes after acknowledgement of the alarm with
“Acknowledge”. The color of the alarm symbol (alarm bell) turns white.
Appendix 227
20.1.2 “Alarm Overview” Menu The alarm overview can be selected as follows:
t Press the “Alarm” function key.
“Alarm Table” Operator Screens
Field Function, entry required

ACK ALL Acknowledges all activated alarms
ACK Acknowledges the selected alarm
RST Resets and deletes the selected alarm
20.2 Process Value Alarms The DCU system has limit value monitoring routines that monitor all process variables
(measured data and calculated process values) to ensure that they are within the
alarm limits (High | Low).
The alarm limits must be within the measurement range limits. After entering the
alarm limits, you can release or lock the limit value monitoring individually for every
process parameter.
The DCU system can lock certain process outputs after process value alarms.
228 Appendix
“Process Value Alarms” Operator Screen

High limit °C Upper alarm limit in the physical unit of the PV
Low limit °C Upper alarm limit in the physical unit of the PV
Alarm Status for alarm monitoring
disabled Alarm monitoring, high | low alarms locked
enabled Alarm monitoring, high | low alarms activated
Appendix 229
20.2.1 Operating Notes Alarms are displayed on the operator screen and must be acknowledged:
Operator Screen Example: Exceeding the Alarm Limit
− If the value falls outside the alarm limits, an alarm window opens above the active
screen. An acoustic signal sounds. The alarm display is displayed in the header line
of the operator screen.
− The process value display also shows a small alarm symbol:
− The alarm window closes after acknowledgement of the alarm with “Acknowledge”
or after pressing .
− After the alarm is confirmed with “Acknowledge”, the alarm symbol disappears.
− After pressing “ ”, the alarm is stored in the alarm list as an unacknowledged
alarm and the alarm symbol remains active (the alarm bell stays red).
− If several alarms have been triggered, the next, still unconfirmed alarm will be
displayed after the active alarm window is closed.
20.2.2 Special Notes The DCU system continues to display limit value alarms as long as the process value
remains outside the alarm limit values.
230 Appendix
20.3 Alarms for Digital Inputs Digital inputs can be prompted in response to alarm conditions as well.
These can be used to monitor components like limit contacts (antifoam | level
sensors), motor protection switches or circuit breakers.
When an alarm is triggered, an alarm message with the time of the alarm event
and an acoustic confirmation signal is emitted.
The DCU system can lock certain process outputs after process value alarms.
“Alarm Monitoring” Operator Screen

Å
Æ
Alarms Param. Alarm Monitoring Operating Mode
disabled Alarm monitoring locked for the input
enabled Alarm monitoring activated for the input
Appendix 231
20.3.1 Operating Notes A new alarm is indicated in two ways:
− When an alarm is triggered for the first time, a message appears in the display and
an acoustic signal is emitted.
− The alarm symbol is displayed in the header line of the operator screen.
t Eliminate the cause of the alarm. Check the function of the component that is
producing the input signal, the corresponding connections, and if necessary the
regulator settings.
t Confirm the alarm with “Acknowledge” or press “X”.
y The alarm window closes.
− After the alarm is confirmed with “Acknowledge”, the alarm symbol disappears
(the alarm bell turns white). The alarm is recorded in the alarm list as a
confirmed alarm (”ACK”).
− After pressing “X”, the alarm is stored in the alarm list as an unacknowledged
alarm and the alarm symbol remains active (the alarm bell stays red).
20.3.2 Special Notes For an overview of alarms that have occurred, you can open the alarm table with the
“Alarm” menu button.
20.4 Alarms, Meaning and

Corrective Measures
20.4.1 Process Alarms The user can switch on and off the individual alarms listed in the following table:
Text in the alarm line Meaning Remedy
[Name] State Alarm Digital input alarm Confirm alarm with “ACK”
[Name] Low Alarm The corresponding process value has fallen Confirm alarm with “ACK”
below its lower alarm limit
[Name] High Alarm The corresponding process value has fallen Confirm alarm with “ACK”
below its lower alarm limit
Jacket Heater Failure Overheating protection in the temperature The tempering system must be refilled
circulation of the double wall has triggered
Motor Failure Overheating protection of the motor responded Allow the motor to cool down
232 Appendix
20.4.2 Process Messages Process messages are displayed in the “Phases” menu [¨ Chapter “18. “Phases”
Menu”]. Both during automatic process control and in single-step control, the header
line on the operator screen shows the process status for the running program, e.g.
“State: Running”.
Text Meaning Remedy

State: Running Sterilization running No intervention required
State: Idle Sterilization program not active Start sterilization with “Start”
Sterilization finished Sterilization is finished The fermentation process can be started by
confirming with “ACK”
20.4.3 System Alarms The alarms in the following table are system-generated messages that the user
cannot switch off:
Text in the alarm line Meaning Remedy

Source: Factory Reset Confirmation message for a system reset Confirm alarm with “ACK”
started from the “Settings” menu
[Name] Watchdog Timeout Confirmation message for a Watchdog Note down the alarm and report it to the
Timeout, triggered by malfunctions in the DCU Service Department.
including reference to the source of failure Confirm alarm with “ACK”
Power Failure Power failure with the date and time Confirm alarm with “ACK”
Power lost at [yyyy-mm-dd
hh:mm:ss]
Power Failure, Process Stopped Power failure with the date and time; Confirm alarm with “ACK”.
System in Standby maximum power outage time exceeded
Power lost at [yyyy-mm-dd
hh:mm:ss]
Shut down Unit # “Emergency off” was pressed on the bioreactor Switch the bioreactor back on with
“Emergency off”
20.5 Bug Handling and If the DCU system should encounter technical problems, contact Sartorius Stedim
Troubleshooting Service.
20.6 Locking Functions Locking functions are permanently configured; the user cannot change them. In the
“Settings” menu, locked inputs and outputs are highlighted with a colored marking.
In the “Phases” menu, locked phases have the status “locked”. The extent of the
locking functionalities is system-specific and is predefined during configuration.
This is documented in the configuration lists enclosed with every system.
20.7 GNU Licensing − DCU systems contain software subject to the license terms of the “GNU General
Public License (GPL)” or the “GNU Lesser General Public License (LGPL)”.
If applicable, the terms of the GPL and LGPL as well as information about the
options for access to GPL code and LGPL code used in this product are available
upon request.
− The GPL code and LGPL code contained in this product are published without any
guarantee and subject to the copyright of one or more authors. You can find
detailed information in the documentation about the enclosed LGPL code and in
the GPL and LGPL terms and conditions.
Appendix 233
20.8 Password System
Only disclose this information to authorized users or service staff. If required, remove
this page from the manual and keep it in a special place.
Certain system functions and settings that should only be accessible for authorized
personnel are protected by the standard password system. These include, in the
regulator menu, the settings for the regulator parameters (e.g. PID), in the “Settings”
menu:
− Process value setting “PV”
− At the manual operation level (”Manual Operation”), the interface parameter
setting for digital and analog process inputs and outputs or for simulation
controllers.
The “Service” submenu of the “Settings” menu is only accessible via a special service
password. This is only provided to authorized service technicians.
When selecting password-protected functions, a key field is displayed automatically

with a prompt to enter the password. The following passwords can be defined:
− Standard password (factory-set: 19)
− Customer-specific standard password1
− Service password1
1 You will receive this information by post or together with the Technical Documentation
234 Appendix
August-Spindler-Str. 11
Phone +49.551.308.0
Fax +49.551.308.3289
www.sartorius-stedim.com
Copyright by
Sartorius Stedim Biotech GmbH,
Göttingen, Germany
No part of this publication may be
reprinted or translated in any form
or by any means without the prior
written permission of Sartorius
Stedim Biotech GmbH.
All rights reserved by Sartorius
Stedim Biotech GmbH in
accordance with copyright laws.
The information and figures

contained in these instructions
correspond to the version date
specified below. Sartorius Stedim
Biotech GmbH reserves the right to
make changes to the technology,
features, specifications and design
of the equipment without notice.
Date:
February 2014
Sartorius Stedim Biotech GmbH,
Goettingen, Germany
Printed in the EU on paper bleached

without chlorine. | W
Publication No.: SBT6022-e140201
Ver. 02 | 2014

Manual Biostat Cplus Sbt6022-E

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Operating Manual

85037-540-72 Vers. 02 | 2014

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Part A: BIOSTAT® Cplus

Part B: DCU4 System for BIOSTAT® Cplus

t If the manual should be lost or misplaced, please contact Sartorius for a

The following presentations will also be used:

− Texts that follow this mark are lists.

y Texts that follow this mark describe the result of an action.

” “ Texts in quotes are references to other chapters or sections.

[¨ ] Texts following this symbol make reference to other chapters, sections or

Sartorius Stedim Systems GmbH

Transport damage, as well as measures for subsequent cleaning and disinfection of

Failure to observe the following safety precautions can have severe

Danger of crushing your hand or fingers in the peristaltic pump – do not

Equipment manufacturers have placed some identifying marks, symbols or labels on

Only biological materials in Groups 1 and 2 may be used in the device.

The intended use also includes:

Danger due to use other than the intended use!

Anyone assigned to the installation, commissioning, operation, maintenance or repair

The device may only be used:

Furthermore, the following rules must be followed:

Non-stationary equipment is any equipment that is designed to be continually moved

2.7 Danger Due to

2.9 Danger Due to Gases

2.9.1 Dangers Due to

Reaction with other materials!

2.9.2 Danger Due to Danger of suffocation due to escaping nitrogen!

2.9.3 Danger Due to

Danger of contamination from escaping feed and culture media!

2.12 Danger Due to

2.14 Danger Due to

Wear the following personal protective

Protective work clothes

2.16.1 Switch-Disconnector Control Unit

Safety Valve / Burst Disk

In addition, there is a safety valve in the temperature control circulation.

The safety valve or burst disk triggers at a defined pressure.

2.17 Emergency Instructions Preventive Measures

In particular, the following apply:

Date Name Type of Training Training Given by Signature

The device is made up of the following components:

The BIOSTAT® Cplus with 5 L culture vessel is a bench-top system.

Overview BIOSTAT® Cplus using the example of the Cplus L (floor-standing)

Fig. 3-3: Control unit connection panel, side

Fig. 3-4: Control unit connection panel, side (continued)

Variable area flow meter

Designations on the variable area flow meter

“O2 Enrichment” Aeration

“Gasflow Ratio” Aeration

Fig. 3-6: Aeration “Advanced Additive Flow - 2 out”

The aeration rates are set at variable area flow meters.

Fig. 3-7: Integrated peristaltic pumps

WM 102 ﬁxed speed 20 rpm

Tubing wall thickness 1.6 mm

* = tubing sizes delivered by default

Culture vessels 10, 15, 20 and 30 liters

Culture vessel 5 Liter

Pos. Description Pos. Description Pos. Description

Fig. 3-8: Cover plate design Cplus 5-2

Pos. Description Placement, mounting, other comments