Operating Manual
UniVessel®
Culture Vessels and Equipment
85032-542-05
Version 02 | 2012
Legal Notices
Figure on title page:
Example of a UniVessel®-type culture vessel
Sartorius Stedim Biotech GmbH
Technical Documentation
August-Spindler-Strasse 11
D-37079 Goettingen
Germany
Website: www.sartorius-stedim.com
E-mail: tech.pubs@sartorius-stedim.com
Printed in Germany on chlorine-free paper.
If you require mandatory information regarding specific
properties of your UniVessel® culture vessel that goes beyond
that which is described here, please contact us.
Reproduction, translation and duplication in any form,
also in part, require written permission from Sartorius
Stedim Biotech GmbH.
UniVessel® is a registered trademark of Sartorius Stedim
Biotech GmbH.
Status: May 2012
Sartorius Stedim Systems GmbH, Goettingen, Germany
Sartorius Stedim Systems GmbH
Robert-Bosch-Str. 5–7
34302 Guxhagen, Germany
Phone +49.5665.407.0
www.sartorius-stedim.com
Copyright by Sartorius Stedim Systems GmbH, Germany.
All rights reserved.
The enclosed CD contains the operating instructions
for UniVessel® as a PDF file in German, French, Italian
and Spanish.
System Requirements:
– Windows, MacOS X
– Browser with JavaScript enabled
– PDF-Reader
Use start.html to run the application
Contents
1.
2.
3.
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Safety Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.8
Overview of the Culture Vessel Program . . . . . . . . . . . . . . . . . . . . 12
1.1 Overview of the Culture Vessel Program . . . . . . . . . . . . . . . . . 12
1.1.1 Vessel Design and Functional Features . . . . . . . . . . . . 12
1.1.2 Culture Vessel Features . . . . . . . . . . . . . . . . . . . . . . . . . 13
1.1.3 Schematic of the Culture Vessel Construction . . . . . . 14
1.2 UniVessel® 0.5 L . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
1.3 UniVessel® 1 L, Jacketed and Single Walled . . . . . . . . . . . . . . . 15
1.3.1 UniVessel® 1 L, Jacketed . . . . . . . . . . . . . . . . . . . . . . . . 15
1.3.2 UniVessel® 1 L, Single Walled . . . . . . . . . . . . . . . . . . . . 15
1.4 UniVessel® 2 L, Jacketed and Single Walled . . . . . . . . . . . . . . . 15
1.4.1 UniVessel® 2 L, Jacketed . . . . . . . . . . . . . . . . . . . . . . . . 15
1.4.2 UniVessel® 2 L, Single Walled . . . . . . . . . . . . . . . . . . . . 16
1.5 UniVessel® 5 L, Jacketed and Single Walled . . . . . . . . . . . . . . . 16
1.5.1 UniVessel® 5 L, Jacketed . . . . . . . . . . . . . . . . . . . . . . . . 16
1.5.2 UniVessel® 5 L, Single Walled . . . . . . . . . . . . . . . . . . . . 16
1.6 UniVessel® 10 L, Jacketed and Single Walled . . . . . . . . . . . . . . 17
1.6.1 UniVessel® 10 L, Jacketed . . . . . . . . . . . . . . . . . . . . . . . 17
1.6.2 UniVessel® 10 L, Single Walled . . . . . . . . . . . . . . . . . . . 17
1.7 Special Vessels and Optional Equipment . . . . . . . . . . . . . . . . . 18
3.9
3.10
3.11
3.12
3.13
Setup, Installation and Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
2.1 Completeness Check . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
2.2 Checking Culture Vessels and Attachments . . . . . . . . . . . . . . . 19
2.3 Workplace . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.3.1 Workplace Setup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.3.2 Tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.3.3 Autoclaving . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.3.4 Dimensions and Space Requirements | Autoclave
Dimensions (Single Walled and Jacketed Vessels) . . . . 22
Internal Equipment of the Vessels . . . . . . . . . . . . . . . . . . . . . . . . . . 24
3.1 Stirrer Shafts and Motor Connections . . . . . . . . . . . . . . . . . . . 24
3.1.1 Stirrer Shafts and Drive Shaft Couplings . . . . . . . . . . . 24
3.1.1.1 Equipment and Specification Drive . . . . . . . 24
3.1.1.2 Operating Information . . . . . . . . . . . . . . . . . 24
3.1.1.3 Installation Information . . . . . . . . . . . . . . . . 25
3.2 Stirrers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
3.2.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 26
3.2.2 Assembly Information . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.2.3 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.3 Baffle Insert and Baffles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
3.3.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 28
3.3.2 Assembly Information . . . . . . . . . . . . . . . . . . . . . . . . . . 29
3.3.3 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 29
3.4 Sparger Pipe with Ring Sparger . . . . . . . . . . . . . . . . . . . . . . . . 30
3.4.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 30
3.4.2 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
3.4.3 Connecting the Gas Supply . . . . . . . . . . . . . . . . . . . . . 31
3.5 Sparger Pipe with Micro Sparger . . . . . . . . . . . . . . . . . . . . . . . 32
3.5.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 32
3.5.2 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.5.3 Connecting the Gas Supply . . . . . . . . . . . . . . . . . . . . . 33
3.6 Sparger Insert . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
3.6.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 34
3.6.2 Installation and Connection . . . . . . . . . . . . . . . . . . . . . 35
3.6.2.1 Connecting the Membrane . . . . . . . . . . . . . . 35
3.6.2.2 Information on Connecting and
Operating the Gas Supply and Exhaust . . . . 36
3.7 Universal Adapter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
3.7.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 37
3.7.2 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
3.14
3.15
3.16
3.17
4.
Inoculation Ports (Septa) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
3.8.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 38
3.8.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 38
Dip Tube for Adding Media or Harvesting . . . . . . . . . . . . . . . . 39
3.9.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 39
3.9.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 40
Spinfilter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
3.10.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 41
3.10.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 43
3.10.2.1 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
3.10.3 Design Examples for Using the Spinfilter . . . . . . . . . . 44
Draft Tube . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
3.11.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 45
3.11.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 46
3.11.2.1 Assembly Inside the Culture Vessel . . . . . . . 46
3.11.2.2 Assembly of Spinfilter and Stirrer . . . . . . . . 46
3.11.2.3 Special Information on In-Process Use . . . . 46
Dummy Plugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
3.12.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 47
3.12.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 47
Quadruple Addition Port Adapter . . . . . . . . . . . . . . . . . . . . . . . 48
3.13.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 48
3.13.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 48
3.13.2.1 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
3.13.2.2 Connection . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Inoculation Ports (Septa) d 19 mm . . . . . . . . . . . . . . . . . . . . . 49
3.14.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 49
3.14.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 49
3.14.2.1 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
3.14.2.2 Transferring the Inoculation Culture
or other Media . . . . . . . . . . . . . . . . . . . . . . . . 49
1-Channel Inoculation Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
3.15.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 50
3.15.2 Operating Instructions . . . . . . . . . . . . . . . . . . . . . . . . . 50
3.15.2.1 Preparing the Inoculation Kit
(Autoclavable Media, Inoculation Culture) . 50
3.15.2.2 Preparing the Inoculation Kit
(Non-autoclavable Media) . . . . . . . . . . . . . . 50
3.15.2.3 Connecting the Inoculation Kit . . . . . . . . . . 51
Antifoam Disc . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
3.16.1 Principle of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . 51
3.16.2 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 51
Port Adapter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
3.17.1 Principle of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . 52
3.17.2 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 52
Electrodes and Sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
4.1 Pt-100 Temperature Sensor . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
4.1.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 53
4.1.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 54
4.1.2.1 Assembly of Electrode Without Core . . . . . . 54
4.1.2.2 Assembly with Core . . . . . . . . . . . . . . . . . . . . 54
4.1.2.3 Operating Notes . . . . . . . . . . . . . . . . . . . . . . 54
4.2 Antifoam and Level Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
4.2.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 55
4.2.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 56
4.3 pH Electrode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
4.3.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 57
4.3.1.1 Reactivating pH Electrodes . . . . . . . . . . . . . . 58
4.3.1.2 Calibrating the pH Electrode . . . . . . . . . . . . 58
4.3.1.3 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
4.3.1.4 Connection and Startup . . . . . . . . . . . . . . . .58
4.3.1.5 Other Notes . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Contents
3
4.4
4.5
4.6
pO2 Electrodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
4.4.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 59
4.4.1.1 Polarographic Electrodes . . . . . . . . . . . . . . . 60
4.4.1.2 Optical pO2 electrodes . . . . . . . . . . . . . . . . . . 60
4.4.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 60
4.4.2.1 Assembly and Connection . . . . . . . . . . . . . . . 60
4.4.2.2 Calibrating Polarographic Electrodes . . . . . 61
4.4.2.3 Calibrating Optical Electrodes . . . . . . . . . . . 61
Redox Electrode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .61
4.5.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 61
4.5.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 61
4.5.2.1 Reactivating Redox Electrodes . . . . . . . . . . . 62
4.5.2.2 Calibrating the Redox Electrode . . . . . . . . . . 61
4.5.2.3 Assembly and Connection . . . . . . . . . . . . . . . 62
Turbidity Electrode FUNDALUX® II . . . . . . . . . . . . . . . . . . . . . . 62
5.
External Accessories for Installation and Connection . . . . . . . . . 63
5.1 Exhaust Cooler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
5.1.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 63
5.1.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 64
5.1.2.1 Assembly on the Culture Vessel
without Extension Kit . . . . . . . . . . . . . . . . . . 64
5.1.2.2 Installation with Extension Kit . . . . . . . . . . . 65
5.1.2.3 Folding up the Exhaust Cooler . . . . . . . . . . . 65
5.1.2.4 Connecting the Cooling Water Supply . . . . 65
5.2 Sterile Filters for Air | Gas Supply and Exhaust . . . . . . . . . . . . 66
5.2.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 66
5.2.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 66
5.2.2.1 Assembling and Connecting
the Inlet Air Filter . . . . . . . . . . . . . . . . . . . . . 66
5.2.2.2 Assembling and Connecting
the Exhaust Filter . . . . . . . . . . . . . . . . . . . . . . 67
5.2.2.3 Exhaust Filter in the Membrane
Aeration Basket . . . . . . . . . . . . . . . . . . . . . . . 67
5.2.2.4 Connecting the Bioreactor . . . . . . . . . . . . . 67
5.3 Cooling Fingers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
5.3.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 68
5.3.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 69
5.3.2.1 Assembly on the Culture Vessel . . . . . . . . . . 69
5.3.2.2 Connecting and Operating the
Cooling Water Supply . . . . . . . . . . . . . . . . . . 69
5.4 STT Quick Connector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
5.4.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 70
5.4.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 71
5.4.2.1 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
5.4.2.2 Connecting the STT Quick Connector . . . . . 71
5.4.2.3 Transferring a Medium . . . . . . . . . . . . . . . . . 71
5.5 Bypass Sampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
5.5.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 72
5.5.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 72
5.6 Other Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
6.
Equipping the Vessel Before Process Start . . . . . . . . . . . . . . . . . . 74
6.1 Corrective Solution Bottles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
6.1.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 74
6.1.2 Operating Information . . . . . . . . . . . . . . . . . . . . . . . . . 74
6.1.2.1 Preparing and Connecting the Bottles . . . . 75
6.1.2.2 Installation in the Peristaltic Pumps . . . . . . 76
6.1.2.3 Pre-adjusting the Corrective Solution
Supply to the Culture Vessel . . . . . . . . . . . . . 77
6.2 Manual Sampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
6.2.1 Equipment and Specifications . . . . . . . . . . . . . . . . . . . 77
6.2.2 Assembly Information . . . . . . . . . . . . . . . . . . . . . . . . . . 78
6.2.3 Operating Notes for Sampling . . . . . . . . . . . . . . . . . . . 79
6.3 Bottle Holder . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
4
Contents
7.
Appendix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
7.1 Cleaning and Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
7.1.1 Measures after Previous Use . . . . . . . . . . . . . . . . . . . 81
7.1.2 Culture Vessel Disassembly . . . . . . . . . . . . . . . . . . . . . 81
7.1.3 Stirrer Shafts and Motor Connections |
Cleaning and Maintenance . . . . . . . . . . . . . . . . . . . . 82
7.1.3.1 Cleaning the Stirrer Shafts after
Process Completion . . . . . . . . . . . . . . . . . . . 82
7.1.4 Stirrers | Cleaning and Maintenance . . . . . . . . . . . . . 83
7.1.5 Baffle Insert and Baffles | Cleaning and
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
7.1.6 Sparger Pipe with Ring Sparger | Cleaning
and Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
7.1.7 Sparger Pipe with Micro Sparger | Cleaning
and Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
7.1.8 Sparger Insert | Inspection, Cleaning and
Maintenance of Culture Vessels . . . . . . . . . . . . . . . . . 84
7.1.9 Universal Adapter | Cleaning and Maintenance . . . . 85
7.1.10 Inoculation Ports (Septa) | Cleaning and
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
7.1.11 Dip Tube for Adding Media or Harvesting |
Cleaning and Maintenance . . . . . . . . . . . . . . . . . . . . 85
7.1.12 Spinfilters| Cleaning and Maintenance . . . . . . . . . . . 86
7.1.13 Draft Tube | Cleaning and Maintenance . . . . . . . . . . 86
7.1.14 Dummy Plugs | Cleaning and Maintenance . . . . . . . 87
7.1.15 Pt-100 Temperature Sensor | Cleaning and
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
7.1.16 Antifoam and Level Probe |
Cleaning and Maintenance . . . . . . . . . . . . . . . . . . . . 87
7.1.17 Quadruple Addition Port Adapter | Cleaning and
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
7.1.18 STT Quick Connector | Cleaning and Maintenance . 88
7.1.19 pH Electrode | Cleaning and Maintenance . . . . . . . . . 88
7.1.20 pO2 Electrodes | Cleaning and Maintenance . . . . . . . 89
7.1.21 Redox Electrode | Cleaning and Maintenance . . . . . 89
7.1.22 FUNDALUX® II | Cleaning and Maintenance . . . . . . . 90
7.1.23 Exhaust Cooler | Cleaning and Maintenance . . . . . . 91
7.1.24 Sterile Filters for Air | Gas Supply and Exhaust |
Cleaning and Maintenance . . . . . . . . . . . . . . . . . . . . 91
7.1.25 Inoculation Ports (Septa) d 19 mm | Cleaning and
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
7.1.26 1-Channel Inoculation Kit | Cleaning
and Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
7.1.27 Cooling Fingers | Cleaning and Maintenance . . . . . . 92
7.1.28 Bypass Sampler | Cleaning and Maintenance . . . . . . 92
7.1.29 Corrective Solution Bottles | Cleaning and
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
7.2 Tempering Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
7.3 Overview of Assembly and Equipment . . . . . . . . . . . . . . . . . . 94
7.3.1 Equipment Options . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
7.3.2 General Instructions on Installing Vessel Equipment 94
7.3.3 Installing New Glass Vessels . . . . . . . . . . . . . . . . . . . 95
7.3.4 Assembly of Glass Vessels on the Stand . . . . . . . . . . . 96
7.3.4.1 Jacketed Culture Vessels . . . . . . . . . . . . . . . 96
7.3.4.2 Single Walled Culture Vessels . . . . . . . . . . . 96
7.3.5 Tubing Connections on Double Walled Glass Vessels 96
7.3.6 Installing Vessel Equipment . . . . . . . . . . . . . . . . . . . . 97
7.3.7 Installation and Connection of Other Equipment . . 97
7.4 Preparing the Culture Vessels for the Process . . . . . . . . . . . . 100
7.4.1 Filling the Jacket of Double Walled Culture Vessels . 100
7.4.2 Filling the Culture Vessels . . . . . . . . . . . . . . . . . . . . . . 100
7.4.3 Other Pre-process Measures to Be Taken . . . . . . . . . 101
7.4.3.1 Mounting the Motor on the Stirrer Shaft . 101
7.4.3.2 Connecting Aeration|Gassing . . . . . . . . . . 101
7.4.3.3 Connecting the Sensors . . . . . . . . . . . . . . . 101
7.4.3.4 Connecting the Corrective Solution
Supply . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
7.4.3.5 Connecting the Tempering System . . . . . . 102
7.4.3.6 Process Start . . . . . . . . . . . . . . . . . . . . . . . . 102
7.5 Assembly and Operation of Special Vessels . . . . . . . . . . . . . . 102
7.6 Decontamination Declaration . . . . . . . . . . . . . . . . . . . . . . . . . 103
Introduction
I. Introduction
Introduction
This document describes the culture vessels and equipment for the bioreactor models
BIOSTAT® Aplus, Bplus, Qplus, B, B-DCU I and B-DCU II supplied by Sartorius Stedim
Systems GmbH.
BIOSTAT® Aplus
BIOSTAT® Bplus
BIOSTAT® B
BIOSTAT® B-DCU I
BIOSTAT® B-DCU II
BIOSTAT® Qplus
The equipment supplied with a bioreactor can include one or more culture vessels
along with the equipment supplied for the reactor type involved and available for the
vessel or may be tailored to the customers’ specifications.
This document illustrates the handling of the culture vessels and equipment –
wherever relevant – how to assemble, install, operate them and how the user can
perform the usual maintenance work on them.
II. Notes on Intended Use
The UniVessel® brand culture vessels are designed for the cultivation of microorganisms and cells in either discontinuous or continuous processes. They may only be
used with equipment and under operating conditions as described in the equipment
manuals or – if the specifications vary – in the supplementary documents.
Modifications to the equipment supplied may only be carried out after consulting with
and obtaining the express consent of Sartorius Stedim Biotech GmbH! Sartorius
Stedim Systems GmbH gives no guarantee and is not liable for use in any other
circumstances (relating to equipment, operating parameters and process sequences).
For safe operation of the culture vessels and equipment, it is required that the users
are qualified and familiar with the handling of the equipment, cultures and media
applied for the intended application, and that they understand the possible hazards
which can occur.
All information provided in this manual refers to the design and equipment of the
culture vessels as current at the time this manual was created and may be subject to
error. The equipment supplied with a bioreactor or culture vessel does not necessarily
include all equipment described herein. The design and specifications of the
components may differ from those described in this manual.
Introduction
5
Notwithstanding its obligations resulting from guidelines, directives, laws and | or
standards governing the interpretation, development and manufacturing or resulting
from contractual agreements, Sartorius Stedim Systems cannot guarantee that the
culture vessels and equipment are suitable for the purpose intended by the user.
The culture vessels and equipment are examples from the product portfolio of
Sartorius Stedim Systems GmbH. If you have any questions about the additional
products in the program, please contact:
Sartorius Stedim Systems GmbH
Robert-Bosch-Str. 5–7
34302 Guxhagen, Germany
Phone +49.5665.407.0
Website: www.sartorius-stedim.com
III. Explanation of Symbols
The “Warning” symbol and this formatting indicate hazards that can be expected
with a high probability and can lead to severe injuries.
The “Caution” symbol and this formatting indicate hazards that can occur and
can lead to severe injuries if the notes and instructions are not observed.
Notes with the “ATTENTION!” warning indicate risks that can lead to material
damage.
Instructions and notes labeled with this symbol indicate steps that must be
performed with particular care or point to other especially important aspects to
be observed.
y
The notice symbol indicates other important information.
1., 2., ...
Numbered paragraphs indicate steps that should be performed consecutively in the
order listed.
t
This symbol points to information referenced in another section.
The use of bioreactors may be connected with potential hazards. These hazards may
result from the energy forms supplied (e.g. electrical power, pressurized media, such
as water, air and gasses), from the devices and equipment themselves (e.g. during
transport, setup or connecting supplies at the workplace) or from decommissioning
and maintenance. The process can also cause further risks, e.g. biological or chemical
risks associated with the cultures or the media used.
The hazard and safety instructions given in this document only apply to the devices
described herein. They supplement the other rules and regulations to be observed
during the process and applicable to the cultures and media employed. Applications
associated with special hazards may mandate the use of additional safety equipment
on the devices or at the workplace and the implementation of special measures to
protect personnel and the work environment. This document does not go into further
detail about such circumstances nor does it deal with regulations legal or otherwise
that may bind the operator or describe the required precautions.
6
Introduction
IV. Safety Information
Organizational Precautions
y Persons responsible for deployment of the equipment must check whether its use
is associated with any special risks to personnel or the work environment. If risks
do exist, appropriate safety provisions must be put in place and safety equipment
must be provided.
y Only authorized persons may stand near the work area and operate devices and
equipment. The personnel must be aware of the potential hazards associated with
the intended process and observe the required precautions to ensure safe
implementation. Safe operation of the devices and equipment mandates that all
users are appropriately qualified and conscientiously familiarize themselves with
their handling.
y Users are required to wear suitable work clothing and appropriate personnel
protective equipment (gloves, safety goggles and a facemask if necessary).
Transport Routes and the Workplace
y Transport routes and the workplace must have the sufficient dimensions and be
able to bear the weight of all devices and equipment. Take the dimensions and the
weight of all parts into account. Use the appropriate tools and assistive devices for
transport and workplace setup.
y The workplace must be suitable to allow cleaning and disinfection of
contaminated equipment or benchtops.
Assembly and Equipment
Danger of injury from shattered glass and glass splinters.
Improper use can damage or break glass vessels (culture vessels, bottles for
corrective solution supply and sampling).
Handle all glass vessels with special care. Check them for any damage.
Never use damaged glass vessels (not even if you only suspect damage).
1. Only use equipment for the culture vessels approved by Sartorius Stedim Systems
GmbH.
2. Check that all vessels and equipment are in perfect working order, especially all
gaskets and O-rings. Replace seals and gaskets whenever they show signs of wear,
e.g. tears or fraying.
Sterilization in the Autoclave
Danger of injury, particularly in the case of large, heavy culture vessels.
Take the weight of the fully equipped and medium-filled culture vessels into
account.
Use the appropriate transportation and lifting equipment.
Accept assistance.
Glass vessels can be destroyed through unallowed overpressure during
autoclaving.
They should not clamp off exhaust air paths. During sterilization, the exhaust
filter ensures sterile pressure compensation between the media space in the
culture vessel and the ambient air.
When using double walled culture vessels, do not close the upper outlet.
The open adapter or tubing connection creates the pressure compensation
between jacket and ambient air.
Safety Information
7
y Before setting up the culture vessel in the autoclave, check that all of the
equipment is assembled properly. All attachment parts and connected peripheral
devices must be carefully and securely fastened together.
y After autoclaving, wait until all the equipment has cooled down.
Wear safety gloves when handling vessels and equipment that are still hot.
In-process Operation
Danger of injury from shattered glass and glass splinters.
Glass vessels (culture vessels, bottles) can break when damaged through improper
use or unallowed overpressurization.
The maximum allowable internal pressure of the culture vessel is 1.3 barg. The
bioreactor controls of the BIOSTAT® series are equipped with a safety valve in the
intake air path. When using other bioreactor controls, the user must ensure that
the internal pressure mentioned above is not exceeded. This can be accomplished
by using an external safety valve.
In the case of double walled culture vessels, the temperature control circulation
must operate without pressure.
Make sure that the water input pressure on the double wall does not exceed
1.5 barg. This can be accomplished by either installing a pressure reduction valve
in the bioreactor control or using an external pressure reducer.
Danger of burns!
Depending on the rpms and the operating time, the stirrer shaft motor can
become very hot.
Never touch the motor with your bare hands.
Wear safety gloves.
Danger of contact with infectious or corrosive substances or other substances
that pose a health risk.
Handle the equipment, cultures, media, extracted samples and the product
carefully. Observe the safety regulations governing cultures, products, residual
substances and contaminated equipment, particularly when infectious or
corrosive media or media that pose a health risk are involved.
Sterilize, disinfect and clean contaminated devices and benchtops thoroughly.
1. Only operate the culture vessels when they are connected to allowable energy
sources, e.g. power connection (motors and heating jackets), cooling water
supplies (temperature control circulation) and gas inlet supplies (air, gasses)
t “PI diagram of the bioreactor”.
2. Only operate the culture vessels within the limits of allowable performance data,
e.g. stirrer’s rpm, temperature, pressure, aeration rates t “Characteristic data,
specifications, operating information”.
3. Avoid contact with rotating parts t “Stirrer shaft motor, couplings”.
4. Monitor the process scrupulously and make sure it is running as intended and
anticipated. Disruptions in the process, malfunctions and defects must not lead to
any unintentional release of cultures and media that could endanger personnel or
work environment or damage devices.
8
Safety Information
Service and Maintenance
Hazard and possible injuries in the case of contact with cultures and media that
pose a health risk.
Always observe the applicable safety guidelines before starting any maintenance
or service work.
When sending (defective) devices and equipment back to Sartorius Stedim
Systems GmbH, you must provided documentation listing the substances they
came into contact with and what kind of decontamination and cleaning
measures were undertaken.
Attach the appropriate Decontamination Declaration to the packaging
t “Sample in the Appendix”.
y Only authorized personnel qualified to do such work may carry out maintenance
and repair work to eliminate faults. If authorized and qualified personnel is not
available, please contact Sartorius Stedim Systems GmbH.
y Only replacement parts that are approved for the culture vessel and the
equipment should be used.
You may not make any technical modifications to devices and equipment unless
Sartorius Stedim Systems GmbH has expressly confirmed that such modifications
do not affect their safe and proper use.
Magnetic Drive for UniVessel®
Danger of injury to people with pacemakers!
The magnetic field emitted from the core of the stirrer shaft can interfere with
or damage equipment which is susceptible to magnetic effects.
Persons with pacemakers must not handle the magnetic drives.
Keep away from devices and objects sensitive to effects of magnetic fields,
such as data storage media and check cards, and do not store them together.
Heating Jackets (for Single Walled Culture Vessels)
Danger of electrical shock in the event of a cable break, faulty heating coils
inside the heating jacket or defective heating coil insulation!
The heating jacket is operated at mains voltage (120/230 V).
Danger of burns! Depending on the operating temperature intended for the
culture vessel, the heating jacket can heat up to approx. 80°C.
After delivery and unpacking, as well as prior to and after any use, visually check
the heating jacket for faultless condition. Never use the heating jacket in the
event that:
– the power cord, foil-shielded heating side, the silicone foam on the insulation
side and the Velcro straps are cracked or porous or show sharp bends or
kinking;
– Black discoloration appears on the heating side, particularly on the power cord
connection. This suggests a defective power cord and | or cable break to the
heating coils in the heating jacket!
Safety Information
9
2
1a
4
3
1b
Possible damage to heating jackets can include:
– Cracks, porosity of the (1a) cable attachment and | or (1b) power cord
– (2) Cracks, porosity of the silicone foil over the heating coils
– (3) Discoloration of the silicone foil (short circuit in heating coil)
– (4) Cracks, porosity of the Velcro fasteners and | or fastening straps
Transport and Assembly
The power cord may be slightly kinked at the connection to the heating jacket
and thereby become damaged.
1. Always lay the heating jacket out flat on the benchtop or lift when rolled out.
Do not roll together more tightly than the jacket is rounded to fit the culture
vessel; do not bend or fold up.
2. Never place objects on the heating jacket. Sharp-edged or heavy objects can
damage the heating coil and cause a short circuit.
3. Never lift or hold the jacket by the power cord, but always grasp it carefully at
the edges opposite the cable connection. During this procedure, the power cord
should be hanging downwards.
4. Wrap the heating jacket around the glass vessel with foil-shielded side touching
the vessel. The side insulated with silicone foam should face outwards and serves
as grip protection. The power cord should be hanging downwards. Carefully insert
the jacket between the stand rods and place them around the glass vessel so that
you can close the Velcro fasteners and | or buckles.
10
Safety Information
Operation
Danger of burns!
When in operation, never touch the heating jacket with your bare hands.
Danger of short circuit or overheating if unallowed voltage is supplied.
Only connect the power cord to the control unit.
This output is controlled through the temperature control on the bioreactor.
Never use any other voltage feed in the lab in accordance with t “Installation
instructions for the bioreactor control”.
1. Lay the power cord so that it cannot be pulled unintentionally.
2. Monitor the heating jacket. Black discolorations occurring on the power cord
connection or along the heating coil indicate that the heating coil and | or cable
are defective. Interrupt operations. Replace the jacket.
3. Whenever there is contact with splashing water or media, interrupt heating
operations, remove the heating jacket from the culture vessel and clean and dry it
thoroughly.
4. In flawless condition, the heating jackets are insensitive to water and the media
used in conventional culture procedures. Test the resistance to the laboratory
acids, alkaline agents and solvents used.
Cleaning, Storage
1. Clean soiled heating jackets carefully using only a wet cloth and warm water or
mild soapy water. Do not use any hard and | or sharp objects or solvents to remove
stubborn soiling.
2. Store the heating jackets clean and dry and do not expose to direct sunlight for
any period of time.
Safety Information
11
1. Overview of the Culture Vessel Program
1. Overview of the Culture Vessel Program
1.1 Overview of the Culture Vessel Program
The UniVessel®-type culture vessels offered by Sartorius Stedim Systems GmbH are
standard equipment on the bioreactors BIOSTAT® Qplus, BIOSTAT® Aplus, BIOSTAT®
Bplus, BIOSTAT® B (equipment supplied from July 1, 2012), BIOSTAT® B-DCU I and
BIOSTAT® B-DCU II. The culture vessels can be supplied as jacketed or single walled
versions. Moreover, special vessels, e.g. airlift and stainless steel vessels, are available.
The product range of individual bioreactors features:
y BIOSTAT® Qplus: UniVessel® with double wall and working volumes of maximum
0.5 L or 1.0 L
y BIOSTAT® Aplus: UniVessel® single walled vessels with working volumes of
maximum 1 L, 2 L and 5 L
y BIOSTAT® Bplus, BIOSTAT® B and B-DCU I: UniVessel® with working volumes of
1 ... 10 L; special versions, such as Airlift vessels 2 L and 5 L, and stainless steel
vessels BS 5 and BS 10 (see separate operating instructions)
y BIOSTAT® B-DCU II: The entire culture vessel program (currently without the
above-mentioned special versions)
1.1.1 Vessel Design and
Functional Features
y Double walled culture vessels are designed for temperature control by means of a
temperature control circulation with water at ambient pressure. To prevent
overpressure from being applied to the double wall, the circulation is connected
to the culture vessel by special quick connect couplings:
– the lower access to the double wall has a self-closing quick connect coupling
– the upper drain from the double wall has an open male tubing adapter
This way, no overpressure can build up during heating from sterilization in the
autoclave
y The single walled vessels are electrically heated with a heating jacket and can be
cooled by a built-in cooling finger. The heating jackets are wrapped around the
glass jacket and controlled by the designated voltage supply powering the supply
unit.
y UniVessel® culture vessels are equipped with impeller drive motors (top-mounted
drive).
The motor can be connected with the stirrer shaft by way of a mechanical or
magnetic coupling.
y UniVessels® have a height to diameter ratio of about 2:1 (or 2.5:1 in the case of
UniVessel® 10 L) and feature the type of concave inner bottom that has proven
effective for culturing microorganisms and animal cells.
y UniVessel® culture vessels are attached to a stand. The holder for the corrective
solution vessels can be fastened to this stand, which facilitates the placement in
the autoclave and ensures a small footprint when set up at the workplace.
y The culture vessels can be autoclaved with their equipment connected, i.e. air inlet
filter, exhaust filter, corrective solution bottles, transfer lines for corrective
solution, accessories for sampling, etc.
The autoclave must be dimensioned large enough to contain the upright culture
vessel fully equipped with its components, but without the motor, see Notes on
t “Dimensions” further down.
y The space required by a bioreactor at the workplace depends on the supply and
control unit, the culture vessels and the peripheral devices connected. You will
find data on the culture vessel dimensions in this manual. Specifications regarding
the space requirements of the bioreactors, e.g. BIOSTAT® Aplus, BIOSTAT® Qplus,
BIOSTAT® Bplus, BIOSTAT® B or BIOSTAT® B-DCU I/II are listed in the respective
bioreactor manuals under the t “Installation” and|or “Installation and Operation”
sections in the technical documentation supplied with each bioreactor.
12
Overview of the Culture Vessel Program
1.1.2 Culture Vessel Features
Standard Equipment:
y Glass vessels featuring a round-bottomed internal medium compartment and
outer double wall for connection to the temperature control circulation or single
walled vessels for operation with an electrical heating jacket
y Polished stainless steel cover with handles, including dummy plugs for all lid ports
y Stainless steel stand with removable holder for storage bottles
y Materials:
– Glass parts made of borosilicate glass
– Product-contact steel parts are made of stainless steel AISI 316 L (1.4435),
the others of stainless steel AISI 304 (1.4301)
– Seals made of EPDM (FDA-approved design)
Equipment options (the actual equipment supplied depends on the bioreactor and the
customer’s specifications)
y Addition | inoculation port with septum
y Multi-way addition ports (3-way for lid ports d 12 mm, 4-way for lid ports
d 19 mm)
y Harvesting pipe with adjustable installation height, harvesting pipe curved for
complete drainage of the culture vessels
y Sterile filters for air | gas supply and exhaust
– Determined by vessel type and size: Midisart membrane filter and | or
Sartofluor filter cartridge
y Exhaust cooler:
y Sensors:
– Pt-100 Temperature sensor
– pH Electrode
– pO2 Electrode
– Antifoam probe
– Level probe
y 3 storage bottles for acid, alkaline, antifoam agent and substrate (250 ml, 500 ml
and 1000 ml)
y Gassing tube with ring sparger or micro sparger
– optional: Aeration basket with silicone membrane insert for bubble-free gas
input
y Insert with baffles
y Stirrer shaft with sliding ring seal
y Direct or magnetic coupling
y 6-blade disc impellers or 3-blade disc impellers
y Manual sampler
y Fundalux turbidity sensors, Redox electrodes
y Inoculation kit M26x1 for lid ports 19 mm
y Bypass sampler
y STT quick connect couplings for sterile connection of external addition or
harvesting vessels
y Spinfilter modules for perfusion operation with
– Filter mesh of different mesh sizes (e.g. 10 μm, 20 μm, 40 μm or 75 μm);
– Special harvesting pipe for removing media from spinfilter module
– Draft tube for vertical loop, if a sparger insert has not been installed
Overview of the Culture Vessel Program
13
1.1.3 Schematic of the Culture
Vessel Construction
This schematic of a double walled UniVessel® is used to explain how the culture vessels
are setup.
Cover
Vessel (double walled)
Holder | stand
Wearing parts kit with hoses (not depicted)
1.2 UniVessel® 0.5 L
Order no.
Description, special features, applications
BB-8846351
UniVessel® 0.5 L, Jacketed
Consists of:
– Double walled glass vessel with round bottom
– Total volume 0.75 L
– Operating volume 0.15 – 0.5 L
– Stainless steel cover
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4031)
14
Overview of the Culture Vessel Program
1.3 UniVessel® 1 L,
Jacketed and Single Walled
1.3.1 UniVessel® 1 L,
Jacketed
Order no.
Description, special features, applications
BB-8846352
UniVessel® 1 L, Jacketed
Consists of:
– Double walled glass vessel with round bottom
– Total volume 1.6 L
– Operating volume 0.4 – 1 L
– Stainless steel cover with vertical handles
– Stainless steel stand for mounting culture vessel
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4301)
1.3.2 UniVessel® 1 L, Single walled
Order no.
Description, special features, applications
BB-8846326
UniVessel® 1 L, Single Walled
Consists of:
– Single walled glass vessel with round bottom
– Total volume 1.6 L
– Operating volume 0.4 – 1 L
– Stainless steel cover with vertical handles
– Stainless steel stand for mounting culture vessel
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4301)
1.4 UniVessel® 2 L,
Jacketed and Single Walled
1.4.1 UniVessel® 2 L, Jacketed
Order no.
Description, special features, applications
BB-8846353
UniVessel® 2 L, Jacketed
Consists of:
– Double walled glass vessel with round bottom
– Total volume 3 L
– Operating volume 0.6 – 2 L
– Stainless steel cover with vertical handles
– Stainless steel stand for mounting culture vessel
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4301)
Overview of the Culture Vessel Program
15
1.4.2 UniVessel® 2 L, single walled
Order no.
Description, special features, applications
BB-8847255
UniVessel® 2L single walled; without accessories
Consists of:
– Single walled glass vessel with round bottom
– Total volume 3 L
– Operating volume 0.6 – 2 L
– Stainless steel cover with vertical handles
– Stainless steel stand for mounting culture vessel
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4301)
1.5 UniVessel® 5 L,
Jacketed and Single Walled
1.5.1 UniVessel® 5 L,
Jacketed
Order no.
Description, special features, applications
BB-8846354
UniVessel® 5 L, Jacketed
Consists of:
– Double walled glass vessel with round bottom
– Total volume 6.6 L
– Operating volume 0.6 – 5 L
– Stainless steel cover with vertical handles
– Stainless steel stand for mounting culture vessel
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4301)
1.5.2 UniVessel® 5 L, Single walled
Order no.
Description, special features, applications
BB-8847278
UniVessel® 5 L, Single Walled
Consists of:
– Single walled glass vessel with round bottom
– Total volume 6.6 L
– Operating volume 0.6 – 5 L
– Stainless steel cover with vertical handles
– Stainless steel stand for mounting culture vessel
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4301)
16
Overview of the Culture Vessel Program
1.6 UniVessel® 10 L,
Jacketed and Single Walled
1.6.1 UniVessel® 10 L,
Jacketed
Order no.
Description, special features, applications
BB-8846355
UniVessel® 10 L, Jacketed
Consists of:
– Double walled glass vessel with round bottom
– Total volume 13 L
– Operating volume 1.5 – 10 L or 5 – 10 L
(depending on the installation length of the equipment)
– Stainless steel cover with vertical handles
– Stainless steel stand for mounting culture vessel
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4301)
1.6.2 UniVessel® 10 L, Single Walled
Order no.
Description, special features, applications
BB-8846356
UniVessel® 10 L single walled, without accessories
Consists of:
– Single walled glass vessel with round bottom
– Total volume 13 L
– Operating volume 1.5 – 10 L or 5 – 10 L
(depending on the installation length of the equipment)
– Stainless steel cover with vertical handles
– Stainless steel stand for mounting culture vessel
Material:
– In contact with product:
– Borosilicate glass
– Stainless steel AISI 316 L (1.4435)
– EPDM (FDA)
– Other:
– Stainless steel AISI 304 (1.4301)
Overview of the Culture Vessel Program
17
1.7 Special Vessels and
Optional Equipment
Documentation on special vessels and optional vessel equipment will only be
supplied if it belongs to equipment supplied with the bioreactor.
A large selection of different customized versions and accessory parts can be
purchased for your specific processes and applications.
If you are interested, have any questions or specific requests, please write to:
info@sartorius-stedim.com
18
Overview of the Culture Vessel Program
2. Setup, Installation and
Startup
2. Setup, Installation and Startup
Before they leave the factory, all culture vessels are put through a comprehensive
function test. This tests covers all components and accessory parts included with the
equipment supplied.
The culture vessels will usually be delivered premounted with the component parts of
the standard equipment or specified in the purchase order t “Order | Delivery
Documents”. Additional components and accessories, such as sensors and accessories
for connecting peripherals, will be packed separately and can be installed or
connected as dictated by the application.
2.1 Completeness Check
You may only equip the culture vessels with components released by Sartorius
Stedim Systems GmbH specifically for that purpose t “Vessel Drawings,
Equipment Lists, Scope of the Order”.
1. Check whether the culture vessels, vessel equipment and accessories have been
delivered in the versions ordered.
2. If any parts of the delivery are wrong or if transport damage has occurred,
please send a claim as soon as possible after receipt.
2.2 Checking Culture Vessels
and Attachments
Danger of injury from shattered glass if glass containers break.
Even hairline cracks can limit their stability to such an extent that safe
operation is no longer possible. Never autoclave or use damaged glass containers
in the process.
1. Before assembling a culture vessel, check to make sure that the components,
particularly all glass parts and seals are not damaged.
2. Verify that all openings and adapters are fitted with proper O-rings and that they
are clean and in proper condition.
Dirty or defective seals can cause contamination. Replace seals regularly.
Slightly lubricate seals with a little silicone grease before fitting the parts.
This can prevent the components from sticking, for example during autoclaving.
Setup, Installation and Startup
19
2.3 Workplace
2.3.1 Workplace Setup
y The culture vessels can be set up at all workplaces, for example on laboratory
benchtops which
– provide sufficient area for the footprint of the entire bioreactor and can bear its
weight
– allow comfortable and safe access to all of the bioreactor’s operational parts
– satisfy the process requirements, e.g. in terms of biological safety, and are
resistant to the effects of the media used, e.g. acids and lyes
– are easy to clean, and, as required, can be disinfected and decontaminated.
y The space requirement depends on the bioreactor, the number and size of the
culture vessels, as well as peripheral equipment to be connected. You will find
dimensional specifications in the drawings of the culture vessels and | or in the
installation instructions included with the bioreactor.
2.3.2 Tools
Each type of work performed on components installed in the UniVessel® described in
the following requires a special set of tools.
To properly carry out all of the work described in this manual, you should
exclusively use the tool kits listed in the table below!
2.3.3 Autoclaving
20
Setup, Installation and Startup
UniVessel® Model
Tool Kit Number
UniVessel® 0.5 L
BB-38247089
UniVessel® 1 L
BB-38247097
UniVessel® 2 L
BB-38247100
UniVessel® 5 L
BB-38247119
UniVessel® 10 L
BB-38247127
Danger of the vessel bursting from overpressurization: Do not clamp off the
exhaust lines for sterilization. During sterilization, the culture vessel is ventilated
through the exhaust filter. Pressure equalization takes place here during and
after sterilization!
Transport the culture vessel and place it in the autoclave in such a way that
previously assembled connection lines and components cannot become loose.
Do not use vacuum autoclaves.
Vacuums can cause the culture medium to foam. Foam penetrating into the inlet
or exhaust filters can cause them to block and render them inoperable.
Autoclave the culture vessel and the connected parts at 121 °C.
Avoid higher sterilization temperatures
This can damage attachment parts, e.g. sensors!
1. For safe sterilization (killing of thermophilic spores), the core temperature in the
culture vessel must be maintained at the sterilization temperature for at least
20 min. As a rule of thumb, sterilization in the 2 L culture vessel takes about
40 min., while that of the 5 L culture vessel takes about 60 min, etc..
2. The conditions for safe sterilization, e.g. the time in the autoclave, must be derived
empirically. Read the documentation on the autoclave. For this purpose, you can
conduct preliminary tests on commercially available test spores (e.g. Bacillus
steathermophilus) in your autoclave.
Before Autoclaving
1. Check all lid ports and installed components. Close any open lid ports. Adapter and
seal screws must be hand-tightened firmly.
2. Place the corrective solution bottles in the holder on the culture vessel. Secure all
movable parts (cables, tubing, hoses, etc.) so that they cannot slip off during
transport. We recommend using cable straps for fastening hoses to the inlets of
the culture vessel.
3. Clamp off the gas inlet tubing and all hoses leading to attachments that are
immersed in the medium. During sterilization, never allow any medium to be
forced into the tubing.
4. The stirrer drive motor cannot be autoclaved. If the motor is already attached to
the culture vessel, unscrew the fastening screw and remove the motor from the
stirrer shaft adapter.
5. Pull the sensor connection cables back as much as possible.
– Wrap aluminum foil around the sensor plugs and the cables of the Pt-100 to
protect them from the direct effects of steam from sterilization.
– Some sensors are supplied with special protective caps that you can attach for
autoclaving. No protection is needed for the antifoam and level probe.
6. Transport the culture vessel with all attached equipment to the autoclave.
Perform sterilization as required to reliably kill all intrusive external germs.
When transporting the equipment to the autoclave and back and when setting it
back up again at the workplace, make sure that none of the connection lines or
components attached to the culture vessel become loose.
After Autoclaving
Danger of burns from equipment that becomes heated during autoclaving.
Allow the vessel and the attached components to cool down in the autoclave.
Wear safety gloves during handling and transport.
1. Check the screw connections on the lids. The thermal expansion created during
heating and cooling may cause them to loosen. Hand-tighten them all again
firmly.
2. Carefully transport the vessel back to the workplace. Place it on the basic unit of
the bioreactor in such a way that enables you to connect all peripheral devices
easily.
Setup, Installation and Startup
21
2.3.4 Dimensions and Space Requirements | Autoclave Dimensions
H 2)
1)
2)
3)
4)
Includes bottle support
With setup exhaust cooler and exhaust filter
With stand, without medium
To lower the height in the autoclave, an adapter is available for the exhaust
cooler which allows it to be folded to one side during autoclaving (see also
t Section 5.1).
W 1)
Fig.: 1–10 L. Single walled vessel
Autoclave dimensions, reduced height 4)
W + H + D in mm
Autoclave dimensions in mm
Weight 3) in kg
Width (W) 1)
Height (H) 2)
Depth (D) 1)
1L
200
540
270
2L
230
600
300
480 + 410 + 300
7
5L
260
730
340
530 + 510 + 410
12
10 L
330
860
420
540 + 630 + 420
19
22
UniVessel®
6
H 2)
W 4)
Fig.: 0.5 L. Double walled vessel
1)
2)
3)
4)
5)
Includes bottle support
With setup exhaust cooler and exhaust filter
With stand, without medium
Includes bottle support and fabric hoses
To lower the height in the autoclave,
an adapter is available for the exhaust cooler
which allows it to be folded to one side
during autoclaving
(see also t Section 5.1).
W 4)
W 4)
Fig.: 1–10 L. Single walled vessel
Autoclave dimensions, reduced height 5) Weight 3) in kg
W + H + D in mm
Autoclave dimensions in mm
Width (W) 4)
Height (H) 2)
Depth (D) 1)
0.5 L (3-compartment tray) 540
350
430
1L
330
540
270
2L
380
600
300
480 + 410 + 300
8
5L
410
730
340
530 + 510 + 410
14
10 L
450
860
420
540 + 630 + 420
23
7
You will find further instructions on assembling the culture vessels in Sections
t “7.3” and t “7.4” of this operating manual.
UniVessel®
23
3. Internal Equipment of the Vessels
3.1 Stirrer Shafts and Motor Connections
3. Internal Equipment of the Vessels
The UniVessel® culture vessels feature stirrer shafts with sliding ring seals made of
silicon carbide | graphite. The stirrer drive motors can be connected to the stirrer shaft
either by a standard coupling with a sliding ring seal (direct drive) or by a magnetic
coupling.
y Stirrer shafts with a sliding ring seal and standard coupling are the proven
standard for cell culture and microbial applications.
y Stirrer shafts with sliding ring seals and magnetic coupling are used when more
stringent requirements are placed on the sterility assurance level (protection
against contamination, e.g. in the case of cell cultures or containment
fermentation processes to prevent the release of infectious media).
y The equipment delivered is stipulated in the purchase order.
3.1.1 Stirrer Shafts and
Drive Shaft Couplings
3.1.1.1 Equipment and
Specifications
3.1.1.2 Operating Information
24
Internal Equipment of the Vessels
A)
1.
2.
3.
4.
Drive coupling between stirrer shaft and direct drive motor
BIOSTAT® Qplus, B-DCU II
UniVessel® 0.5 L
UniVessel® 1–5 L
BIOSTAT® Aplus
UniVessel® 1–10 L
BIOSTAT® Bplus, B, B-DCU I/II
UniVessel® 1 L
BIOSTAT® Qplus
BB-8841231
BB-39240983
BB-8847320
BB-8841221
B) Magnetic coupling between stirrer shaft and motor
UniVessel® 1 L–10 L
BIOSTAT® Bplus, B-DCU I/II
BIOSTAT® B
UniVessel® 1 L–10 L
BB-8847339
BB-8840359
C)
1.
2.
3.
4.
5.
BB-8841213
BB-8846340
BB-8847347
BB-8847355
BB-8847363
Stirrer shaft with single sliding ring seal
UniVessel® 0.5 L
BIOSTAT® Qplus, B-DCU II
UniVessel® 1 L
UniVessel® 2 L
UniVessel® 5 L
UniVessel® 10 L
You will find instructions on how to mount the motor in the corresponding Section
of this manual. Instructions on how to configure the stirrer’s rpms are given in the
t “Installation Manual for the Bioreactor”.
3.1.1.3 Installation Information
Disassembly, Maintenance and Installation (Mechanical Coupling)
1
Coupling half D30/10H7
Coupling (stirrer shaft part)
BB-38348012
3
4
Set screw
Set screw
BB-39504689
V-ring
V-seal
BB-39122476
2
7
Ring D36-32/11
Ring
BB-38351501
O-ring 33.05 x 1.78
EPDM | FDA BB-0001379
5
1. Take the cover plate off the culture vessel and place it on a flat benchtop.
2. Remove the stirrers from the shaft.
3. Turn the bearing housing counterclockwise and unscrew the stirrer shaft out of
the middle lid port.
4. Check the O-Ring (5). If soiled, clean or replace it if porous surfaces or pressure
points are identifiable.
5. Insert the stirrer shaft into the middle lid port. Turn the bearing housing clockwise
and screw the stirrer shaft on tightly.
6. Attach the stirrers you require for the intended process.
7. Attach all other equipment to the cover plate that has to be screwed on tightly
from underneath.
8. Attach the cover plate to the culture vessel.
You will find instructions on how to clean the stirrer shaft under
t “7.1.3 Stirrer Shafts and Motor Connections | Cleaning and
Maintenance”.
O-ring 29.82x2.62
EPDM | FDA BB-39121780
2–10 L with mechanical sliding ring seal
1
2
6
3
7
4
5
8
9
11
10
Disassembly, Maintenance and Installation (Magnetic Coupling)
Stirrer shafts with magnetic coupling are supplied either premounted or together with
a retrofitting kit which allows users to exchange the direct coupling to a magnetic one
themselves.
1. Remove the stirrer shaft part of the coupling from the standard drive coupling (11).
In doing so, leave the ring and V-seal attached to the shaft!
2. Attach the half of the permanent magnetic coupling (5) to the stirrer shaft such
that the V-seal (9) is under slight pressure.
3. Using set screw (8), fasten the half of the permanent magnetic coupling (5) to the
stirrer shaft.
4. Insert the matching piece to the permanent magnetic coupling (4) into the
adapter sleeve (3).
5. Using set screw (2), fasten the adapter (1) to the motor shaft and insert it into the
adapter sleeve (3).
6. Insert the adapter (1) with motor into the adapter sleeve (3) and fasten it using
the matching permanent magnetic coupling (4) and set screw (7).
7. Now insert the adapter (1) with motor, adapter sleeve (3), and permanent
magnetic coupling (4) onto the stirrer shaft (10) and fasten it with the knurled
screw (6).
You will find instructions on how to clean the stirrer shaft under
t “7.1.3 Stirrer Shafts and Motor Connections | Cleaning and
Maintenance”.
Internal Equipment of the Vessels
25
3.2 Stirrers
Culture vessels designed for microbial cultures (”MO” versions) include 6-blade disc
impellers. These stirrers are especially suitable for intensive mixing of the culture at
high stirring speeds, e.g. to provide high oxygen inputs.
The culture vessels designed for cell cultures (”CC” versions) include 3-blade disc
impellers. They allow for gentle but effective mixing of the culture medium even at
low stirring speeds and are suited for cells sensitive to shearing forces. When used
together with a tubing aeration system, they circulate the medium in a vertical loop.
If a spinfilter is installed, optimum flow against the filter mesh is achieved.
Paddle stirrers are particularly suited for cultivating adherent cells. They prevent the
sedimentation from microcarriers forming on the vessel bottom.
All stirrers are made of stainless steel AISI 316 L (1.4435). The angles of the 3-blade
disc impellers are adjustable such that users can set both the angle as well as the
mixing direction (from top to bottom or bottom to top along the stirrer shaft)
themselves.
3.2.1 Equipment and
Specifications
A) 6-blade disk impeller
1. UniVessel® 0.5 L
2. UniVessel® 1 L
3. UniVessel® 2 L
4. UniVessel® 5 L
5. UniVessel® 10 L
Stirrer diameter 40 mm
Stirrer diameter 45 mm
Stirrer diameter 53 mm
Stirrer diameter 64 mm
Stirrer diameter 75 mm
BB-8841214
BB-8846359
BB-8846804
BB-8847371
BB-8847380
B) 3-blade segment impeller (standard stirrer)
1.) UniVessel® 0.5 L
Stirrer diameter 40 mm
2.) UniVessel® 1 L
Stirrer diameter 48 mm
3.) UniVessel® 2 L
Stirrer diameter 54 mm
4.) UniVessel® 5 L
Stirrer diameter 65 mm
5.) UniVessel® 10 L
Stirrer diameter 78 mm
BB-8841227
BB-8846367
BB-8847398
BB-8847401
BB-8847410
C) Paddle stirrers
1.) UniVessel® 1 L
2.) UniVessel® 2 L
3.) UniVessel® 5 L
4.) UniVessel® 10 L
BB-8848640
BB-8848641
BB-8848642
BB-8848643
Stirrer diameter 48 mm
Stirrer diameter 54 mm
Stirrer diameter 65 mm
Stirrer diameter 78 mm
Compatibility of existing stirrers from other culture vessels:
Stirrers are especially adapted to the dimensions of the culture vessels and of the
stirrer shaft. When installed correctly, they cannot hit other equipment installed
in the vessel.
Stirrers of existing culture vessels may be used if they can be securely fastened to
the stirrer shaft. They must not hit against other internal equipment of the
vessels. Even at high stirring speeds, the stirrers must not release unintentionally.
26
Internal Equipment of the Vessels
3.2.2 Assembly Information
y The height at which the upper stirrer must be attached depends on the filling level
of the vessel. The lower stirrer must be fastened approx. 1 cm above the ring or
micro sparger.
y When using the bladed segment impeller, note the rotating direction of the stirrer
shaft. The vertical flow can be directed down or up depending on the alignment of
the impeller blades and when applied together with an aeration basket or a draft
tube.
1. Remove the cover plate from the culture vessel.
2. Loosen the screws (4) to adjust the height on the stirrer shaft.
Slide the stirrers to their desired height.
3. For fitting other stirrers, remove the existing stirrers from the stirrer shaft, attach
the new stirrers and slide them into the desired position.
y The equipment supplied with “CC” type vessels includes a bladed disc impeller.
If you need additional stirrers (6*), you must purchase them separately.
4. Carefully fasten the stirrers using the screws (4). The stirrers must be secured
against loosing even at high stirring speeds or with highly viscous culture media.
How to install various stirrer types:
(2) 6-blade disc impellers ("MO” vessels)
(6) Bladed segment impeller ("CC” vessels).
The figure shows 2 of the 3 stirrer blades
3.2.3 Operating Information
Stirrer type, stirrer arrangement, angle of segment stirrer blades and stirring speed
should be coordinated with the other equipment installed inside the vessel so as to
achieve optimum turbulence and mixing of the culture medium.
Check to make sure that no dead zones occur in the medium compartment at low
stirring speeds or that fluid vortices are not generated at high stirring speeds.
Otherwise, the exchange of substances taking place in the culture medium and
the supply of microorganisms and cells with oxygen and | or nutrients can be
impaired. Avoid such stirring speeds or change the arrangement of the attachment parts in the culture vessels so as to minimize these effects.
y The arrangement of disc impellers when used in conjunction with a tubing
aeration system, draft tube or spinfilter is illustrated in the corresponding sections
of this manual.
Internal Equipment of the Vessels
27
3.3 Baffle Insert and Baffles
Baffle inserts provide flow resistance to achieve better mixing of the culture medium
even at low stirring speeds and to improve mass transfer and oxygen input. At high
stirring speeds, they help prevent the generation of fluid vortices.
The baffle insert on UniVessels® 1 L, 2 L and 5 L consists of a ring-shaped insert with
4 baffles. On the UniVessel® 10 L, the equipment comprises a single baffle along with
a sparger pipe to which an additional baffle is attached (not depicted).
3.3.1 Equipment and
Specifications
28
Internal Equipment of the Vessels
BB-8846375
Baffle insert UniVessel® 1 L
y d OD = 106.5 mm, HB = 112 mm, 4 baffle inserts, Htot = 153 mm
BB-8846812
Baffle insert for UniVessel® 2 L
y d OD = 125 mm, HB = 130 mm, 4 baffle inserts,
Htot = 200 mm
BB-8846820
Baffle insert for UniVessel® 5 L
y d OD = 154 mm, HB = 204 mm, 4 baffle inserts, Htot = 300 mm
BB-8846839
Baffle for UniVessel® 10 L
y Single baffle HB = 280 mm, Htot = 433 mm
(the gas inlet pipe carries an additional baffle)
y Installed in the lid port d 6 mm opposite the gas inlet pipe
3.3.2 Assembly Information
Adapter
Observe the arrangement in your vessel or follow the specifications in the drawing on
the lid. Alternatively, you can choose a lid port with the appropriate distance to the
stirrer shaft.
1. Remove the cover plate.
O-ring
Extension rod
Baffle insert
2. Unscrew the hexagon nut and pull the baffle insert and | or the baffle out of the
lid port.
3. Check the O-ring and replace if soiled or damaged.
4. To install, push the adapter from underneath up through the lid port.
Baffles
5. Center the baffle insert in relation to the stirrer shaft.
y On the UniVessel® 10 L, a baffle is attached to the air inlet pipe. You can mount
the additional baffle in a lid port opposite the air inlet pipe. Align the separate
baffle towards the center of the vessel.
6. Screw the hexagon nut from above and tighten carefully.
Assembly of the baffle insert
3.3.3 Operating Information
y For mounting an aeration basket or a draft tube for vertical loop, dismantle the
baffle insert | the single baffle.
y On UniVessel® 10 L, one baffle is attached to the air inlet pipe. It is possible to
mount additional baffles into free lid ports d 6 mm opposite to or at the same
angular distances. Direct the baffles either towards the stirrer shaft and | or
towards the center of the vessel.
Internal Equipment of the Vessels
29
3.4 Sparger Pipe with Ring Sparger
The sparger pipe with ring sparger is particularly suitable for intensive gassing in
microbial fermentations. The ring sparger features evenly distributed fine holes
through which the air or gas can bubble into the culture medium.
The sparger pipe with ring sparger is standard equipment on “MO” type culture vessels.
In other vessel types, it can be installed instead of other gassing devices (micro
spargers, sparger inserts) for processes that require intensive gassing, where bubbling
does not affect cell growth and the culture medium does not tend to foam.
3.4.1 Equipment and
Specifications
30
Internal Equipment of the Vessels
BB-8841223
Sparger pipe for UniVessel® 0.5 L
y Sparger pipe with ring sparger, for lid port 6 mm
y Insertion depth 126 mm
y Hose connector for gas supply tubing d ID = 3.2 mm
y Stainless steel AISI 316 L (1.4435)
BB-8846383
Sparger pipe for UniVessel® 1 L
y Like sparger pipe for UniVessel® 0.5 L, here with
insertion depth 170 mm
BB-8846847
Sparger pipe for UniVessel® 2 L
y Like sparger pipe for UniVessel® 0.5 L, here with
insertion depth 229 mm
BB-8846855
Sparger pipe for UniVessel® 5 L
y Like sparger pipe for UniVessel® 0.5 L, here with
insertion depth 336 mm
BB-8846863
Sparger pipe for UniVessel® 10 L
y Like sparger pipe for UniVessel® 0.5 L, here with
insertion depth 453 mm
y with baffles
3.4.2 Assembly
y The sparger pipe is usually located in the lid port as shown in the drawing of the
cover plate or in a free lid port d 6 mm.
y The cover plate must be dismantled from the culture vessel.
1. Unscrew the hexagon nut (3) and pull the sparger pipe down and out of the lid
port.
2. Lubricate the new O-ring with a little silicone grease. This will prevent it from
sticking in the lid port.
3. To install, push the adapter (1) from below up into the lid port.
Center the ring sparger in relation to the stirrer shaft. Screw in the hexagon nut
(3) from above and hand-tighten firmly.
y The insertion depth HE is dictated by the adapter (1). When using culture vessels
with agitators, the lowermost stirrers should be positioned approx. 1 cm above the
ring sparger.
Sparger Pipe with Ring Sparger
3.4.3 Connecting the Gas Supply
1. Attach the hose connected to the exhaust filter when preparing the culture vessel
for sterilization.
2. Before sterilization, clamp off the hose with a hose clamp. That way, no medium
can be forced into the sparger pipe during sterilization. This would block the holes
in the ring sparger and hinder the flow of air | gas into the medium.
3. To set up the culture vessel at the workplace after the autoclaving and prepare it
for the process, connect the hose between air inlet filter and gas supply on the
bioreactor.
4. Remove the hose clamp between air inlet filter and sparger pipe when you want
to start with gassing.
Internal Equipment of the Vessels
31
3.5 Sparger Pipe with
Micro Sparger
Micro spargers are suited for gassing cells sensitive to shearing forces in culture media
that only have a slight tendency to foaming. The micro sparger consists of a frit made
of sintered stainless steel. Air or gas can be introduced into the culture medium
through the pores of the frit to create small bubbles.
The sparger pipe with micro sparger is standard equipment on “CC” type culture
vessels. It can be ordered as an optional accessory for other culture vessels.
It is installed instead of the existing gassing device (i.e. the sparger pipe with ring
sparger or aeration basket).
3.5.1 Equipment and
Specifications
32
Internal Equipment of the Vessels
BB-8841215
Gassing pipe with micro sparger 500 mL
y Gassing pipe with gassing frit for installing into 6 mm lid port
y Insertion depth 160 mm
y Gassing frit with 10 μm pores
y Hose connector for tubing with d ID = 3.2 mm
Materials:
y Frit stainless steel AISI 430 F (1.4404), pipe stainless steel
AISI 316 L (1.4435)
BB-8846391
Gassing pipe with micro sparger 1 L
y Like micro sparger 0.5 L, insertion depth 170 mm
BB-8807760
Gassing pipe with micro sparger 2 L
y Frit micro sparger 0.5 L, insertion depth 225 mm
BB-8807779
Gassing pipe with micro sparger 5 L
y Like micro sparger 0.5 L, insertion depth 332 mm
BB-8807787
Gassing pipe with micro sparger 10 L
y Like micro sparger 0.5 L, insertion depth 455 mm
3.5.2 Assembly
Attach the sparger pipe in such a way that air and | or gas cannot bubble out of
the frit directly onto the pO2 electrode. This would impair the pO2 measurement.
y The sparger pipe is usually located in the lid port as shown in the drawing of the
cover plate or in a free lid port d 6 mm.
1. Take the cover plate off the culture vessel.
2. Unscrew the hexagon nut (3) and pull the sparger pipe downwards out of the
lid port.
3. Lubricate the new O-ring (2) with silicone grease. This will prevent it from sticking
in the lid port.
4. Push the adapter (1) from below up into the lid port. Line up the frit (4). Screw on
the hexagon nut (3) from above and tighten carefully.
y The insertion depth HE is dictated by the length of the sparger pipe.
The lowermost stirrer should be positioned approx. 1 cm higher than the frit.
Sparger pipe with micro sparger
3.5.3 Connecting the Gas Supply
1. Attach the hose connected to the exhaust filter when preparing the culture vessel
for sterilization.
2. Before sterilization, clamp off the hose with a hose clamp. That way, no medium
can be forced into the sparger pipe during sterilization. This would clog up the
holes in the ring sparger and prevent the air or gas input into the medium.
3. To set up the culture vessel at the workplace after the autoclaving and prepare it
for the process, connect the hose between air inlet filter and gas supply on the
bioreactor.
4. Remove the hose clamp between air inlet filter and sparger pipe when you want
to start with gassing.
Internal Equipment of the Vessel
33
3.6 Aeration Basket
Not available for UniVessel® 500 ml.
Aeration basket with silicone membranes are culture vessel attachments designed for
bubble-free aeration of the culture medium by means of gas diffusion across a silicone
membrane. They are used instead of sparger pipes with ring sparger and | or micro
sparger used, particularly for gassing tissue cell cultures.
Gas inlets that supply air, air enriched with O2 (or pure oxygen), nitrogen, or CO2 for
acidic pH control can be connected to the aeration basket. Which gasses and how
they are supplied (pulsed or at continuously controlled flow) depends on how the
bioreactor is equipped.
The silicone membrane provides a reversible diffusion of O2, N2 and CO2.
When there is excess gas in the medium, the gasses also diffuse back into the
membrane and into the exhaust air. The differential partial pressure of a gas in the
gaseous and | or liquid phase determines the direction and intensity of the gas
exchange.
The silicone membrane insert is optional equipment for culture vessels. If included in
the equipment ordered, the silicone membrane will be preinstalled in the vessel. It can
also be purchased separately and alternatively installed in existing aeration systems.
Gas supply devices for UniVessel®:
– Aeration basket with silicone membrane
– Sparger pipe with ring sparger
– Sparger pipe with micro sparger
3.6.1 Equipment and
Specifications
BB-8846405
BB-8848009
BB-8848017
BB-8848025
34
Internal Equipment of the Vessels
Silicone membrane insert UniVessel® 1 L for bubble-free aeration of
cell cultures
y Install in 6 mm lid port
Consists of:
y Holder for silicone membrane, made of stainless steel AISI 316 L
(1.4435)
y Silicone membrane, length 2.7 m, diameter 3 mm,
wall thickness 0.35 mm
y 2 adapters 6 mm (inlet and exhaust), 2 membrane filters for inlet
and exhaust, pressure controller
y 1+ 3-blade segment impeller
Aeration basket, silicone membrane, diameter 3 mm,
wall thickness 0.35 mm for
UniVessel® 2 L, length 5.2 m
UniVessel® 5 L, length 10.4 m
UniVessel® 10 L, length 16 m
y Mount the stirrers (3) on the stirrer shaft. If necessary, determine the following in
advance:
– The insertion height for optimum circulation around the aeration basket.
– The insertion angle of the stirrer blades that achieves the desired circulation.
3.6.2 Installation and Connection
y If an aeration basket is used in addition to an optional spinfilter, the spinfilter
must be installed first. Follow the instructions given.
3.6.2.1 Connecting the Membrane
1. Insert the holding rod (9) into the cover plate and screw it tight.
2. Mount 2 hose nozzles for the gas supply and the gas outlet on the cover plate.
Use the lid ports as indicated in the t “Vessel lid drawings”.
3. Connect the hose nozzles on the aeration basket to the hose nozzles in the cover
plate using 2 pieces of silicone hose 3.2 +1.6. Secure the tubing connections
against inadvertent slipping.
4. For the head space ventilation, you can install a t “universal adapter” in the
cover plate.
5. After installing the other accessories, mount the cover plate on the culture vessel.
(2) Stirrer shaft
(3) Bladed segment impeller
(5) Holder
(6) Aeration membrane
(7) Dip tube
(8) Foam probe
(9) Holder for aeration basket
9
8
7
2
6
5
3
Connecting the aeration basket
Internal Equipment of the Vessels
35
3.6.2.2 Information on
Connecting and Operating
the Gas Supply and Exhaust
Overpressure can cause the silicone membrane to burst! The internal pressure in
the membrane must not exceed 1.3 barg.
The aeration membrane should always be immersed in the culture medium to
achieve optimum gas diffusion exchange. If an additional spinfilter is installed,
the culture medium must not flush into the spinfilter. Keep the filling level of the
vessel between the upper edge of the membrane insert and the spinfilter.
1. Connect the inlet and exhaust filter to the hose nozzles in the cover plate.
2. For optimum gas diffusion, the internal pressure of the silicone membrane must
be precisely adjusted. For this purpose, attach the pressure hold unit (12) to one of
the stand rods on the culture vessel and connect the exhaust line to the pressure
controller. After activating the gassing, you can then adjust the internal pressure
on the exhaust side.
3. When filling the culture vessel with medium ensure that the aeration membrane
is fully covered with medium. When taking samples from the culture vessel, the
filling level should not exceed the upper edge of the aeration basket. Add more
medium under sterile conditions.
Outlet gassing tubing
Outlet gassing tubing
Gas supply
Gas supply
Exhaust
External connections of aeration basket:
(1) Glass vessel
(2) ... (9) See figure on previous page
(10) Gas inlet filter
(11) Exhaust filter
(12) Pressure controller
36
Internal Equipment of the Vessels
Exhaust
Complete installation with spinfilter (13)
t “Description of Spinfilter"
3.7 Universal Adapter
The universal adapter allows you to set up lid ports for connecting gas inlet or exhaust
from the aeration basket and from the culture vessel headspace. You can also use the
adapter for the corrective solution supply, for instance if all other access options to
the culture vessel are occupied, e.g. the quadruple addition port t Description
“Quadruple addition ports”.
3.7.1 Equipment and
Specifications
BB-8807809
Universal adapter 6 mm
y For connecting corrective solution inlet supply lines as well as
gas inlet and outlet lines on the tubing aeration system or for
deaeration of the headspace
y Install in 6 mm lid port
y Hose nozzle (outside of the vessel) d OD = 6 mm
y Hose nozzle (inside of the vessel) d ID = 4 mm
y Made of stainless steel AISI 316 L (1.4435)
BB-8807817
Universal adapter 8 mm
y For deaeration of the headspace and connecting inlet lines for
corrective solution
y Assembly, materials, same as universal adapter 6 mm
y Hose nozzle (outside of the vessel) d OD = 8.5 mm
3.7.2 Assembly
y The cover plate must be dismantled from the culture vessel.
1. Insert the adapter (1) into a lid port d = 6 mm from below and screw it in tight
with the hexagon nut (2) from above.
2. For connecting the aeration basket, you need 2 adapters. Connect the connection
tubing leading to and from the aeration basket at the lower connector
t “Description of the aeration basket”.
3. When connecting a corrective solution supply, connect the hose (3.2+1.6) to the
upper hose nozzle t “Description for connecting corrective solution supply”.
4. For deaeration of the headspace, connect, a sterile filter to the upper hose nozzle
via a short piece of silicone hose t “Description of the sterile filters”.
5. Secure all hoses with hose clamps.
Universal adapter 6 mm
Internal Equipment of the Vessels
37
3.8 Inoculation Ports (Septa)
Inoculation ports (septa) enable you to spike the culture vessel in-process without the
danger of contamination and to introduce the inoculation culture or special media.
The self-sealing inoculation membrane can be pierced repeatedly. You can introduce
the media using an injection syringe with sterile needle, for example.
3.8.1 Equipment and
Specifications
BB-8844631
3.8.2 Operating Information
Assembly
y The cover plate must be dismantled from the culture vessel.
Cap
O-ring, cap
Sleeve
Membrane
Nut
Septum port kit 6 mm
y For piercing with an injection syringe or thin inoculation needle
y Install in 6 mm lid port
y Made of stainless steel AISI 316 L (1.4435)
1. Push the adapter into the cover plate from below and screw it in tight from above
with the hexagon nut.
2. Place the new inoculation membrane in the adapter. Screw on the sleeve and
hand-tighten firmly.
3. Place the cap over the sleeve.
Adapter
y The cap protects the membrane against contamination until you need to pierce
the inoculation port for transferring inoculation culture and | or corrective
solution.
O-ring, adapter
Fitting the Septum Port Kit B | MD
Pierce with a injection syringe for inoculation or media transfer
1. Remove the seed material or the medium from the storage container using a
sterile injection syringe. While protecting it against contamination, bring the
syringe to the culture vessel.
2. Remove the protective cap from the inoculation port. Flame the membrane.
When using disinfecting agent, allow time for it to work.
3. Pierce the inoculation membrane with the syringe and transfer the seed material.
Afterwards, take the syringe back out, disinfect the membrane once more and
replace the protective cap.
38
Internal Equipment of the Vessels
3.9 Dip Tube for Adding Media
or Harvesting
Dip tubes can be used for the submersed addition of media.
y You can use a dip tube for adding or harvesting media to take samples from a
holding tank, to connect the manual sampler t “Manual sampler” or a bypass
sampler t “Bypass sampler”.
y When used for removal in a holding tank, you can press the medium out of the
culture vessel by applying slight overpressure through the dip tube or suction it
off using a pump. You can automate the process of removing media and pumping
with the level controller t “Level controller” in the manual on the measurement
and control system].
y You can connect a STT quick coupling to the sampling tubing. This gives you the
option of connecting multiple vessels in sterile series t Description of “STT quick
connect coupling”.
y You can use non-sterile or sterile sampling or harvesting bottles as collection
vessels.
3.9.1 Equipment and
Specifications
There are two types of dip tubes (also referred to as sampling, addition or harvesting
pipes); all dip tubes are inserted into the 6 mm d lid port:
1. Height adjustable.
BB-8841228
Dip tube 0.5 L
BB-8848637
Dip tube 1 L
BB-8807884
Dip tube 2 L
BB-8807892
Dip tube 5 L
BB-8807906
Dip tube 10 L
2. With curved endpiece for complete drainage.
BB-8848632
Dip tube 0.5 L
BB-8848633
Dip tube 1 L
BB-8848634
Dip tube 2 L
BB-8848635
Dip tube 5 L
BB-8848636
Dip tube 10 L
Internal Equipment of the Vessels
39
3.9.2 Operating Information
Assembly
y The cover plate must be dismantled from the culture vessel.
Dip tube with curved end pipe (Fig. A):
y Insert the dip tube into the lid port from below and screw the adapter (2) on
tightly with the hexagon nut (4).
Dip tube with adjustable insertion height (Fig. B):
1. Insert adapter (2) for dip tube into the lid port and screw on tightly with the
hexagon nut (4).
A
2. Insert clamping cone (5) and screw cap (6) onto the dip tube.
3. Insert dip tube into the adapter and push down to the desired immersion depth.
y The maximum immersion depth is achieved when the dip tube is advanced until
it hits the locking ring (7).
4. Screw the dip tube on tight. The clamping ring (5) secures the pipe against
slipping.
B
Dip tube assembly:
Fig. [A]: Fixed dip tube
Fig. [B]: Height-adjustable dip tube
After sterilization, never advance the adjustable dip tube deeper into the vessel.
This may cause contamination.
40
Internal Equipment of the Vessels
Connecting transfer line and sample vessels
1. Attach the tubing for sample transfer and secure against slipping. Consider the
required hose length when installing the hose in a pump.
2. If you autoclave the sampling container together with the culture vessel, you can
connect the hose to it directly.
3. If you are using the STT quick coupling, connect the hose to the coupling
connector t “STT quick connect coupling"; when using the manual sampler,
connect the hose to the inlet connector t “Manual sampler”.
4. For sterilization in the autoclave, clamp off the hose to prevent medium from
being driven out of the culture vessel.
Sampling | Product Harvesting
1. If not yet connected to a sampling container, place the hose in a holding tank.
When using the STT quick coupling, attach the STT male connector of the inlet line
to the sample container. When using a manual sampler, follow the description
given.
2. Open the hose clamp. Frequently, the marginal overpressure generated by
aeration in the culture vessel and the force of gravity are sufficient to transfer
the sample. You can clamp off the exhaust line briefly. This will raise the pressure
in the culture vessel and forces the sample out.
y For sampling via the level controller, place the hose in a peristaltic pump.
3. Clamp the hose off again after sampling. To prevent contamination when you
pump samples into a vessel openly, immerse the free end of the tubing in a beaker
containing disinfecting agent in between two sampling steps.
3.10 Spinfilter
Not available for UniVessel® 0.5 L and 1 L.
Cannot be used with Airlift vessels.
3.10.1 Equipment
and Specifications
Spinfilters are used in cell cultures for removing media from the culture vessel.
The sleeve consists of a 4-layer square mesh.
The different mesh sizes allow you to determine the filter behavior in relation to the
cell sizes and the desired retention or separation of cell fragments.
For example, spinfilters are suited for retaining microcarrier-bound cells, while
allowing suspended cells and | or cell fragments to be removed.
In order to ensure problem-free operation, the spinfilter should always be operated
in combination with an aeration basket or a draft tube (see next section). This is the
only way to ensure vertical loop flow around the spinfilter.
Internal Equipment of the Vessels
41
BB-8847447
BB-8808309
BB-8808317
BB-8808325
Spinfilter made of stainless steel
BB-8847448
BB-8847452
BB-8847460
BB-8847487
BB-8847449
Disposable spinfilter
BB-8847991
BB-8847479
BB-8847495
BB-8808260
BB-8808279
BB-8840598
Spinfilter 10 μm, UniVessel® 2L
y Installation on stirrer shaft
y 4-layer square mesh made of stainless steel AISI 430 F (1.4404),
pore size 10 μm
Spinfilter 20 μm, UniVessel® 2 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 2 L
Spinfilter 40 μm, UniVessel® 2 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 2 L
Spinfilter 75 μm, UniVessel® 2 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 2 L
Spinfilter 10 μm, UniVessel® 5L
y Installation on stirrer shaft
y 4-layer square mesh made of stainless steel AISI 430 F (1.4404),
pore size 10 μm
Spinfilter 20 μm, UniVessel® 5 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 5 L
Spinfilter 40 μm, UniVessel® 5 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 5 L
Spinfilter 75 μm, UniVessel® 5 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 5 L
Spinfilter 10 μm, UniVessel® 10L
y Installation on stirrer shaft
y 4-layer square mesh made of stainless steel AISI 430 F (1.4404),
pore size 10 μm
Spinfilter 20 μm, UniVessel® 10 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 10 L
Spinfilter 40 μm, UniVessel® 10 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 10 L
Spinfilter 75 μm, UniVessel® 10 L
y Setup and equipment same as spinfilter 10 μm, UniVessel® 10 L
Dip tube spinfilter 2 L
y Removing media from the filtrate chambers of the spinfilter,
insertion depth HE = 147 mm
y Pipe 4+1 for silicone hose 3.2 +1.6 mm
Dip tube spinfilter 5 L
y Same as dip tube spinfilter 2 L, insertion depth HE = 228 mm
Dip tube spinfilter 10 L
y Pipe 6+1, insertion depth HE = 336 mm
Disposable spinfilters are available on request.
42
Internal Equipment of the Vessels
3.10.2 Operating Information
The dip tubes SF (7) should be installed in a lid port located near the stirrer shaft in
such a way that allows you to draw off the medium from the inside the spinfilter.
3.10.2.1 Assembly
The spinfilter (6) must be attached to the stirrer shaft so that the upper rim is always
kept above the liquid level (the culture medium must not flow over the rim).
y Before installing the spinfilter, take a volumetric measurement of the intended
operating volume and label the filling level on the culture vessel.
1. Remove the existing stirrers from the shaft.
2. Mount the dip tube SF (7) into one of the inner lid ports.
y The dip tubes on the UniVessel® 5 L and 10 L have an angular pipe.
They can be turned to extend sufficiently into the spinfilter.
3. Place the spinfilter (6) onto the stirrer shaft and slide it upwards.
The adjusted height results from the max. filling level of the culture medium.
The medium must not overflow.
4. Secure the spinfilter to the stirrer shaft with the screw (5).
5. Mount the disc impeller onto the stirrer shaft.
6. Finish installing the vessel equipment and mount the cover plate back on.
7. Connect the dip tube SF to the harvesting hose.
Assembly of the spinfilter:
(2) Stirrer shaft
(3) Bladed segment impeller
(4) O-ring
(5) Fixing screw
(6) Spinfilter
(7) Dip Tube SF
Internal Equipment of the Vessels
43
3.10.3 Design Examples for
Using the Spinfilter
y Left-hand figure: Showing installation of the spinfilter in a 2 L culture vessel as
used for cultivating of microcarrier-bound cells
y Right-hand figure: Showing installation of the spinfilter together with an
aeration basket as used for tissue cell cultures
Outlet
gassing tubing
Spinfilter in standard vessel:
(1) Glass culture vessel
(2) Stirrer shaft
(3) Bladed segment impeller (overlapping)
(5) Fixing screw
(6) Spinfilter
(7) Dip Tube SF
(8) Level probe
44
Internal Equipment of the Vessels
Gas supply
Installation with a sparger insert:
(2) Stirrer shaft
(3) Bladed segment impeller (overlapping)
(5) Sparger insert
(6) Aeration membrane
(7) Dip tube SF
(8) Level probe
(9) Holder for sparger insert
(10) Connection to air inlet filter
(11) Connection to exhaust filter
(13) Spinfilter
3.11 Draft Tube
The draft tube is used for cultures where you want to achieve vertical loop mixing of
the medium in the culture vessel. A typical application is the perfusion operation with
a spinfilter when you are not using an aeration basket. The draft tube is applied
together with a bladed segment impeller. At a given rotating direction of the stirrer
shaft, a vertical loop results from the angle of adjustment of the stirrer blades:
1. Inside the draft tube up t and down between draft tube and inner vessel wall.
2. Inside the draft tube down t and up between draft tube and inner vessel wall.
If the draft tube is used in combination with a spinfilter, you will have to test whether
the circulation shown in (1) improves harvesting or whether the spinfilter mesh is
slightly blocked. If the draft tube is used without a spinfilter, the circulation (2)
achieves good distribution of the air (gaseous) input into the culture medium by the
ring sparger|micro sparger, even at low stirrer speeds. When you want to cultivate
microcarrier bound cells, you can check whether this form of circulation is more
favorable.
3.11.1 Equipment and
Specifications
BB-8840539
BB-8840660
BB-8840580
Draft tube 2 L
y Installation in the culture vessel instead of the baffle insert
y Insertion height HE = 200 mm,
– Dimensions HL+d = 115 +76.1 mm,
– Stainless steel AISI 316 L (1.4435)
Draft tube 5 L
y Installation, material same as draft tube 2 L, equipped with 2 lid
ports
y Insertion height HE = 309 mm, dimensions HL+d = 180 +104 mm
Draft tube 10 L
y Installation, material same as draft tube 2 L, equipped with 2 lid
ports
y Insertion height HE = 434 mm, dimensions HL+d = 262 +129 mm
Configure dummy plugs in accordance with t “Vessel lid drawings”!
Ordering information on the draft tube can be found in its description!
Internal Equipment of the Vessels
45
3.11.2 Operating Information
3.11.2.1 Assembly Inside the Culture Vessel
y To install the draft tube in UniVessel® 1 L and 2 L you need 1 lid port, to install
UniVessel® 5 L and 10 L you need 2 lid ports.
y Select the lid port(s) at a distance to the stirrer shaft which allows for centering
the draft tube around the stirrer shaft.
UniVessel® 5 l, 10 l
1. For maintenance and rebuilding of the draft tube, dismantle the cover plate.
O-ring
2. Unscrew the hexagon nut(s) and pull the draft tube out of the lid port(s).
3. Check the O-rings and replace if contaminated or damaged.
4. To install, insert the adapter(s) (1) through the lid port (s) from below.
5. Align the stirrer shaft so that it is in the center of the draft tube.
6. Screw the hexagon nut(s) from above and tighten them carefully.
Assembly of the draft tube
3.11.2.2 Assembly of Spinfilter
and Stirrer
1. Mount the spinfilter on the stirrer shaft in such a way that its upper edge extends
over the draft tube. The culture medium must be able to flow around the draft
tube, but must not flow into the spinfilter when the spinfilter is open, even during
intensive stirring.
2. Mount the bladed segment impeller on the stirrer shaft. Depending on the
circulation intended for the draft tube, you can fasten the stirrers inside or
underneath the draft tube.
3.11.2.3 Special Information on
In-Process Use
1. Keep the filling level of the culture medium between the upper rim of the draft
tube and that of the spinfilter.
2. If you are not using an automatic level controller, you must check the filling level
in the culture vessel when inputting nutrient solutions and corrective solution,
as well as after each sampling step, and then add or remove medium as necessary.
3. If you are using an automatic level controller, set the insertion height of the level
probe to this maximum filling level.
y The measurement and control system offers a variety of methods for level control
t Documentation on the measurement and control system. For perfusion
operation using a spinfilter, you can compensate for the harvested filtrate, e.g. by
automatically adding fresh medium.
y On some bioreactors, an automatic level control can also be achieved by means of
weight-based control. See t Gravimetric flowcontroller in the documentation
accompanying the measurement and control system.
46
Internal Equipment of the Vessels
3.12 Dummy Plugs
Insert dummy plugs into every lid port that is not filled with an accessory part.
y In the case of lid ports d 6 mm, before attaching the cover plate to the glass
vessel you should decide on which openings (ports) you need for the vessel.
When these lid ports are used, it is no longer possible to remove dummy plugs
d 11 and retrofit with other attachment parts once the cover plate has been
assembled.
3.12.1 Equipment and
Specifications
BB-8807957
BB-8807949
BB-8807930
Dummy plugs d 11
y for lid port d = 6 mm
Dummy plugs PG 13.5
y for lid port d = 12 mm
Dummy plugs d 19
y for lid port d = 19 mm
In the case of customer-specified culture vessels, the vessel equipment may vary!
3.12.2 Operating Information
Dummy plugs 6 mm
It is recommended that, when lid ports are not required for attaching accessories
parts, you use them as inoculation ports or plug up them with universal adapters.
This allows in-process access to the vessel, e.g. if you want to introduce medium
additives or nutrient solutions. Only insert dummy plugs in the lid ports that you
truly do not intend to use.
Assembly
1. Insert dummy plugs d 11 through the lid port from underneath and screw them
tight from above with the hexagon nut.
2. In the case of lid ports d = 12 mm, first attach the appropriate gasket. Screw the
dummy plugs 12 mm (PG 13.5) on tight from above.
O-ring
3. You can screw dummy plugs d = 19 mm (M26 +1) on from above and carefully
hand tighten.
Dummy plugs 12 mm
Gasket
After sterilization in the autoclave:
Check to make sure that dummy plugs 12 mm and 19 mm are still screwed on
tightly.
Dummy plugs 19 mm
O-ring
Dummy plugs for lid ports
d 6 mm, d 12 mm and d 19 mm
Internal Equipment of the Vessels
47
3.13 Quadruple Addition
Port Adapter d 19 mm
The quadruple addition port adapter allows you to install four connections for adding
corrective solutions in one lid port. To install the adapter you need one free lid port
d 19 mm.
3.13.1 Equipment and
Specifications
BB-8844313
Quadruple Addition Port
y Install in d 19 mm lid port with thread M 26x1
y 4 integrated tube connectors for tubing of d ID = 1.6 mm
y Made of stainless steel AISI 316 L (1.4435)
3.13.2 Operating Information
1. Screw the adapter (1) into a lid port d 19 mm and hand-tighten it carefully.
3.13.2.1 Assembly
2. After sterilization in the autoclave, verify its proper seat.
3.13.2.2 Connection
1. Connect the tubing to the corrective solution supplies to the hose nozzles (2).
2. Secure all hoses with hose clamps.
2
1
3
Quadruple addition ports
48
UniVessel®
In the case that there are hose nozzles that you do not connect to any inlets, you
can insert a piece of silicone hose in them and clamp them off or knot the tubing.
3.14 Inoculation Ports (Septa)
d 19 mm
The inoculation port d 19 mm allows you to spike the culture vessel without danger of
contamination when you want to add larger volumes of seed culture or additional
media than is possible through the t standard septum. To install it, the culture vessel
must have one free lid port d 19 mm.
You can attach 1-channel inoculation kit. The inoculation membrane is self-sealing
and can be pierced repeatedly for sterile operation (e.g. under flame protection).
3.14.1 Equipment and
Specifications
BB-8840601
Septum port kit d 19 mm
y Install in d 19 mm lid port with thread M 26x1 +1
y Made of stainless steel AISI 316 L (1.4435)
y for 1-channel inoculation kit, order no. BB-8840610
3.14.2 Operating Information
3.14.2.1 Assembly
1. Place the inoculation membrane (2) in the lid port (1).
2. Screw the membrane holder (3) into the lid port to fasten the membrane.
3. Screw the dummy plug (4) into the membrane holder (3). It protects the
membrane against contamination until you connect the inoculation kit.
O-ring
Inoculation port d 1
3.14.2.2 Transferring the Inoculation Culture or Other Media
You can transfer the seed culture or the media using an injection syringe or connect
an inoculation kit (as described under t “1-Channel Inoculation Kit“).
1. Unscrew the dummy plug (6) from the membrane holder. You can flame the
membrane, e.g. with a Bunsen burner. When using disinfecting agent, allow time
for it to work.
2. Pierce the syringe or inoculation needle through the inoculation membrane.
For controlled transfer of the medium you can install a peristaltic pump in the
transfer line.
3. You can leave the inoculation needle in the septum for as long as necessary. If you
take the syringe or inoculation needle back out again after transferring the seed
material or medium, flame the membrane once more and screw the dummy plug
back on.
UniVessel®
49
3.15 1-Channel Inoculation Kit
The 1-channel inoculation kit allows you to pierce a culture vessel in-process without
the risk of contamination, e.g. to add seed culture, special media or corrective solution.
Its special advantage is that it allows you to transfer larger volumes quickly. To connect the inoculation kit, you need one lid port with a inoculation port d 19 mm
t Section “Inoculation Port (Septa) d 19 mm”.
3.15.1 Equipment and
Specifications
BB-8840610
1-Channel inoculation kit
y For connecting silicone hoses 3.2 +1.6 mm to inoculation ports
d 19 mm
y Inoculation needle d 6 +0.5 mm (3) with adapter, screw threads
M 26+1
y 2 m silicone hose 3.2 +1.6
y Equipment supplied including sterile sleeve
y Made of stainless steel AISI 316 L (1.4435)
3.15.2 Operating Instructions
3.15.2.1 Preparing the Inoculation Kit (Autoclavable Media,
Inoculation Culture)
1. Attach the silicone hose for the connection to the reservoir bottle or seed culture
container onto the hose nozzle (4). Cut the hose to the required length and
connect it.
2. Screw on the sterile sleeve (5). This will keep the inoculation needle sterile until
you need the inoculation kit for inoculation and transfer of the seed culture or
other media.
3. Sterilize the inoculation kit with the transfer hose and the connected corrective
solution bottle and | or the seed culture container.
4. Inoculate the container for the seed culture and cultivate it until it reaches the
intended state for transfer to the culture vessel.
3.15.2.2 Preparing the Inoculation Kit (Autoclavable Media,
Inoculation Culture)
1. If you want to filter in non-autoclavable media under sterile conditions, mount
a sterile filter in the tubing line leading to the inoculation kit. Sterilize the
inoculation kit including line and filter.
2. Connect the line to the reservoir bottle when transferring the medium.
50
UniVessel®
3.15.2.3 Connecting the
Inoculation Kit
1. Remove the dummy plug (6) from the membrane holder (7). Flame the membrane
briefly or spray with a suitable disinfecting agent.
The disinfecting agent can enter into the vessel and destroy the culture.
2. Remove the sterile sleeve (5) from the inoculation kit (1). Flame the inoculation
needle briefly and pierce it (3) perpendicularly through the membrane (6) under
flame protection.
3. Screw on the adapter with the cap nut (2) firmly in the membrane holder (7).
4. Transfer the seed culture or medium.
5. Afterwards, you can leave the inoculation needle in the membrane holder (7) and
the hose clamped off.
6. If you want to remove the inoculation kit to connect another line later:
– Remove the inoculation needle from the membrane
– Flame the needle and inoculation port
– Flame the dummy plug or new inoculation needle and screw them in
Connecting the Inoculation Kit
3.16 Antifoam Disc
3.16.1 Principle of Operation
Frequently, foaming in the medium can occur during fermentation processes. This can
have diverse and unintentional side effects, for example, it can block the exhaust
filter.
The antifoam disc counteracts foaming by mechanically destroying the foam. This
takes place directly where the foam is generated, i.e. on the surface of the medium.
The rising foam gets caught by the rotating antifoam disc. There, the rotational
motion separates the gaseous from the fluid components. The gaseous components
escape through the exhaust air path | exhaust filter, while the fluid components are
returned to the medium.
3.16.2 Equipment and
Specifications
Application range (UniVessel® sizes):
1–10 L
Dimensions (disc |shaft):
74 |10 mm – UniVessel 1 L, BB-8844465
84 |10 mm – UniVessel 2 L, BB-8844466
96 |14 mm – UniVessel 5 L, BB-8844467
96 |16 mm – UniVessel 10 L, BB-8844468
Material:
y Polyetheretherketone (PEEK)
Antifoam disc
Required rotating speed of the stirrer:
6 min., 300 rpm
Mounting: y To the stirrer shaft
Mounting position:
1 –1– 3 mm above the filling level (medium)
Sterilization: y Autoclavable
UniVessel®
51
3.17 Port Adapter 19 mm | 2 mm
3.17.1 Principle of Operation
3.17.2 Equipment and
Specifications
52
UniVessel®
This adapter reduces a 19 mm port down to a 12 mm port and is inserted into the
cover plate for this purpose.
BB-8848630
Adapter 19 mm -> 12 mm
4. Electrodes and Sensors
4. Electrodes and Sensors
4.1 Pt-100 Temperature Sensor
4.1.1 Equipment and
Specifications
BB-33197016
Pt-100 for UniVessel® 0.5 L, BIOSTAT® Qplus
y Insertion depth HE = 113 mm
BB-33197024
Pt-100 for UniVessel® 1 L, BIOSTAT® Bplus, Qplus
y Insertion depth HE = 151 mm
Pt-100, type 200-4, for UniVessel® 2 L, BIOSTAT® Bplus
y Insertion depth HE = 211 mm
Pt-100, type 300-4, for UniVessel® 5 | 10 L, BIOSTAT® Bplus
y Insertion depth HE = 316 mm
Pt-100, type 400-4, for UniVessel® 10 L, BIOSTAT® Bplus
y Insertion depth HE = 418 mm
BB-33197032
BB-33197040
BB-33197024
BB-33197105
BB-33197083
BB-33197091
BB-8848647
BB-8848648
BB-8848649
BB-8848650
BB-8848651
Pt-100 with open port for UniVessel® 1 L, BIOSTAT® Aplus
y Insertion depth HE = 151 mm
Pt-100 with open port for UniVessel® 2 L, BIOSTAT® Aplus
y Insertion depth HE = 211 mm
Pt-100 with open port for UniVessel® 5 L, BIOSTAT® Aplus
y Insertion depth HE = 316 mm
Pt-100 for UniVessel® 0.5 L, BIOSTAT® B-DCU II
y Insertion depth HE = 113 mm
Pt-100 for UniVessel® 1 L, BIOSTAT® B, B-DCU II
y Insertion depth HE = 151 mm
Pt-100 for UniVessel® 2 L, BIOSTAT® B, B-DCU II
y Insertion depth HE = 211 mm
Pt-100 for UniVessel® 5 | 10 L (min. AV 5 L), BIOSTAT® B, B-DCU II
y Insertion depth HE = 316 mm
Pt-100 for UniVessel® 10 L (min. AV 1.5 L), BIOSTAT® B, B-DCU II
y Insertion depth HE = 418 mm
Electrodes and Sensors
53
4.1.2 Operating Information
Moisture in the signal cable and plug of the Pt-100 can disrupt the temperature
measurement signal. It has been observed in older sensors that, despite being
sealed, negative pressure can cause condensation to penetrate during
sterilization in the autoclave.
Pt-100 sensors can be installed directly into the fitting in the cover plate.
To prevent moisture from penetrating, you can use the optional open port.
Before sterilization in the autoclave, first only install the open port in the
culture vessel and place the Pt-100 in the open port after sterilization in the
autoclave.
4.1.2.1 Assembly of Electrode
Without Core
y Normally, the Pt-100 temperature sensor is mounted in the lid port as indicated in
the drawing for the culture vessel t“Recommended arrangement of accessory
parts”.
1. To remove a Pt-100, to install the core and | or to check the O-rings in the
adapter, unscrew the cap nut on the Pt-100 and pull it out of the fitting.
Disassemble the cover plate. Then, you can remove the cap nut from the fitting
and pull it out from the lid port.
2. Insert fitting into the cover plate from below and screw tight with the hexagon
nut.
3. After assembling the cover plate, push the Pt-100 into the fitting from above
and screw tight with the cap nut. The length of the sensor shaft dictates the
immersion depth HE in the culture vessel.
4. Before sterilization in the autoclave, wrap aluminum foil around the plug to
protect it from the direct effects of steam.
5. In the autoclave, the plug with the contacts should be hanging downwards so that
no condensation can collect at the contacts.
4.1.2.2 Assembly with Core
1. After disassembling the standard fittings, insert the core into the cover plate from
below.
2. Tightly screw the hexagon nut from the outer side onto the core.
3. After sterilization in the autoclave, install the Pt-100.
During autoclaving, attach the protective cap onto the open port to prevent
condensation from collecting.
4.1.2.3 Operating Notes
y After sterilization in the autoclave, plug the connecting cable of the Pt-100 into
the measurement amplifier panel of the measurement and control systems.
y Do not calibrate the Pt-100.
54
Electrodes and Sensors
4.2 Antifoam and Level Probe
Antifoam and level probes are combination electrodes that work according to the
conductivity principle. The stainless steel parts on the culture vessel act as a counter
electrode.
When foam or medium comes into contact with the probes, it causes a jump in
conductivity. This current jump in the measuring signal activates of the designated
controller.
The sensors are insulated with a ceramic sheath. This sheathing minimizes the risk of
short circuiting the conductivity signal in the event that there is overgrowth of cells
or deposits from media components, etc. The antifoam probe has a defined insertion
height. The fitting for the level probe allows you to vary the insertion height.
4.2.1 Equipment and
Specifications
Level probes:
BB-8844488
BB-8844490
BB-8844690
BB-8844491
Level probe 0.5/1/2/ 5 L, complete, for BIOSTAT® Aplus, Bplus,
B-DCU, Qplus
y Install with fitting in 6 mm lid port
y Probe length 150 mm, insertion height HE = max. 122 mm,
height adjustable
Level probe 1/2/5 L, complete, for BIOSTAT® B, B-DCU II
y Install with fitting in 6 mm lid port
y Probe length 150 mm, insertion height HE = max. 122 mm,
height adjustable
Level probe 10 L, complete, for BIOSTAT® Aplus, Bplus, B-DCU
For 19 mm lid port
y Probe length 300 mm, insertion height HE = max. 260 mm,
height adjustable
y Working volume: UniVessel® 10 L 1,5 ... 10 L
Level probe 10 L, complete, for BIOSTAT® B, B-DCU II
For 19 mm lid port
y Probe length 300 mm, insertion height HE = max. 260 mm,
height adjustable
y Working volume: UniVessel® 10 L 1,5 ... 10 L
Antifoam probes:
BB-8844461
Antifoam probe, complete, for BIOSTAT® Aplus, Bplus, B-DCU
and Qplus
y Install in 6 mm lid port
y Probe length: 80 mm
BB-8844463
Antifoam probe for BIOSTAT® B, B-DCU II
y Install in 6 mm lid port
y Probe length: 80 mm
Connecting cable for individual sensors, probes and electrodes:
(The cables are included in the above-mentioned probe sets for Aplus, Bplus,
Qplus and B-DCU I)
BB-8848684
BB-8848683
Cotter pin|M12 for level probe BIOSTAT® B, B-DCU II
Cotter pin|M12 for antifoam probe BIOSTAT® B, B-DCU II
Electrodes and Sensors
55
y The insertion height is dictated by the installation length of the probe and screw
connection. The insertion height of the antifoam probe is fixed, the insertion
height of the level probe depends on the operating volume in the culture vessel.
4.2.2 Operating Information
y Existing filling volume marks on culture vessels give approximate readings.
The actual filling volume for a certain filling level depends on the accessories
installed.
Determining the Insertion Height
y The insertion height of the antifoam probe is dictated by the shaft length.
Connection cable
y The insertion height of the level probe depends on the the filling volume that
should not be exceeded. For the standard probe, the max. insertion height is
122 mm; for the probe with extended shaft, it is 260 mm below the underside
of the lid.
y Important note: High stirring speeds and intensive gassing raise the fluid level.
Electrode
Insert electrode
from above
1. Measure the filling level of the intended filling volumes on the culture vessel and
the corresponding insertion height.
Cap nut
2. Fasten the probes at the desired height.
Hexagon nut
Assembly
1. Insert the probe adapter in the lid port from below. Screw the adapter tight from
above with the hexagon nut.
O-ring
2. After assembling the cover plate, insert the probe in the adapter from above.
Adapter
Install from
above
3. Slide the probe up to the insertion height and fasten it there with the cap nut.
Antifoam and | or level probe
Connecting the Electrodes
1. The probe cables cannot be autoclaved. If the cables are connected, remove them
before placing the culture vessel in the autoclave.
2. The inner O-ring in the adapter provides a sterile seal for the level probe. In case
the probe is set too deep, it is possible to pull it out a bit after sterilization. After
sterilization, never slide the probe deeper into the vessel. This can introduce germs
and thereby cause contamination.
3. After autoclaving and|or before process start, plug the cables into the appropriate
“Foam” and|or “Level” sockets on the control unit.
When using bioreactors with multiple culture vessels (BIOSTAT® Bplus Twin, Qplus,
B-DCU), note which measurement amplifier panel is assigned to the respective
culture vessel.
y Instructions on configuring the operating parameters are included in the
documentation t “Measurement and control system”.
4.3 pH Electrode
56
Electrodes and Sensors
Sterilizable combination electrodes with pasteous bridging electrolyte are used to
measure the pH. The advantages of these sensors over conventional combination
electrodes filled with liquid electrolytes are:
Gel-filled pH electrodes are pressurized at the factory. They can work in environments
at up to 2.5 barg pressure (max. allowable internal pressure for UniVessel® is 1.3 barg).
The sensors are maintenance-free. Although a small amount of the electrolyte always
diffuses through diaphragm here as well, the electrolyte lasts for the whole lifetime of
the electrode.
Hamilton sensors are in standard use by Sartorius Stedim Biotech.
4.3.1 Equipment and
Specifications
BB-34090810
BB-34090811
BB-34090812
BB-34090813
BB-34090814
BB-8848654
BB-8848655
BB-8848656
BB-8848657
BB-8848658
pH Electrode EF—12/120 K8-HM UniVessel® 0.5 L, for BIOSTAT® Qplus
With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 120 mm
pH Electrode EF-12/160/2 K8-HM UniVessel® 1 L, for BIOSTAT® Aplus,
Bplus, Qplus
With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 160 mm
pH Electrode EF-12/200 K8-HM UniVessel® 2 L, for BIOSTAT® Aplus,
Bplus
With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 200 mm
pH Electrode EF-12/325K8-HM UniVessel® 5/10 L (5 L min. AV),
for BIOSTAT® Aplus, Bplus
With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 325 mm
pH Electrode EF-12/425 K8-HM UniVessel® 10 L (1.5 L min. AV),
for BIOSTAT® Bplus
With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 425 mm
pH Electrode, VP-HM – UniVessel® 0.5 L, BIOSTAT® B-DCU II
y With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 120 mm
pH Electrode, VP-HM – UniVessel® 1 L, BIOSTAT® B, B-DCU II
y With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 160 mm
pH Electrode, VP-HM – UniVessel® 2 L, BIOSTAT® B, B-DCU II
y With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 225 mm
pH Electrode, VP-HM – UniVessel® 5 L and 10 L (5 L min. AV),
BIOSTAT® B, B-DCU II
y With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 325 mm
pH Electrode, VP-HM – UniVessel® 10 L (1.5 L min. AV),
BIOSTAT® B, B-DCU II
y With internally pressurized reference electrolyte gel (Hamilton)
y Electrode length 425 mm
pH electrodes from Mettler-Toledo are available on request.
Connecting cable for individual sensors, probes and electrodes:
(The cables are included in the above-mentioned probe sets for Aplus, Bplus,
Qplus and B-DCU I)
BB-8848681
VP8|VP8 for pH sensor BIOSTAT® B, B-DCU II
Calibration buffer for pH electrodes:
BB-34090366
PH 9, 21 Hamilton, 250 ml
BB-34090367
PH 4
Hamilton, 250 ml
BB-34090368
PH 7
Hamilton, 250 ml
Electrodes and Sensors
57
4.3.1.1 Reactivating pH
Electrodes
4.3.1.2 Calibrating the pH
Electrode
Operating Information
1. Before first use or after it has dried out after long-term storage or after dry
storage, you must carefully reactivate your pH electrode.
Glass membranes, as used in pH electrodes, form a thin aqueous gel layer that
determines the measuring properties (response time, slope, alkaline error etc.).
This layer needs to regenerate.
2. Follow the instructions in the t “Manufacturer’s documentation”. You can place
your pH electrode for approx. 10 min in 0.1–1M NaOH , then in 0.1–1M HCl and
for around another 15 min. in the manufacturer’s storage solution.
Calibration buffers can cause caustic burns!
Caution is advised when handling buffer solutions. Wear safety gloves.
y You must calibrate the zero point and the slope of the pH electrodes prior to
installation in the culture vessel t Documentation on the measurement and
control system, manufacturer’s documentation on the pH electrode.
For calibration, use the buffer solutions supplied with the electrode
(also available as consumables).
4.3.1.3 Assembly
Danger of damaging the pH electrode.
Other attachment parts in the culture vessel, particularly the rotating stirrers,
may not butt up against the electrode. Use a suitable lid port, e.g. one located
further away.
1. Insert the pH electrode into a free lid port d 12 mm.
O-ring
The insertion depth Hso depends on the culture vessel and shaft length. For 10 L
vessels, there are sensors with different lengths, depending on the minimum operating
volumes of 5 L or 1.5 L.
2. Screw in the pH electrode fitting hand tight. Never use a pliers or wrench!
3. To prevent the pH electrode from drying out, fill the culture vessel with water or
culture medium after you have installed all parts.
Installing the pH Electrode
4.3.1.4 Connection and Startup
1. The cable must not be autoclaved. To protect the sensor plug from the effects of
steam during autoclaving, you can attach the protective cap supplied or wrap
aluminum foil around the plug.
2. After autoclaving and setting up the culture vessel at the workplace and|or before
starting the process, connect the electrode on the measurement and control
system. When using bioreactors with multiple culture vessels (BIOSTAT® Bplus
Twin, Qplus, B-DCU), note which measurement amplifier panel is assigned to the
respective culture vessel.
3. Configure the pH measurement and control parameters t Documentation on the
measurement and control system.
58
Electrodes and Sensors
4.3.1.5 Other Notes
y In order to be able to account the potential effects caused by heat or the culture
medium during autoclaving on measuring behavior of the pH electrode, the pH
electrode can be recalibrated in-process t “Recalibration function /
Recalibration, measurement and control system”.
4.4 pO2 Electrodes
The dissolved oxygen is measured as partial oxygen pressure pO2 using sterilizable pO2
electrodes. The pO2 electrodes operate on the polarographic principle. They consist of
an Ag anode and Pt cathode, which are separated by a gas-permeable polymer
membrane. An electrolyte with defined layer thickness between membrane and
cathode conductively connects anode and cathode. At a suitable polarization voltage,
the oxygen diffusing through the membrane will be reduced completely at the
cathode. This chemical reaction produces an electrical current in the mA-range, which
is directly proportional to the partial oxygen pressure.
The permeability of the membrane depends on temperature. At rising temperature the
electrode current will rise by around 3% per degree °C. This temperature effect is
compensated by a thermistor integrated in the electrode.
4.4.1 Equipment and
Specifications
4.4.1.1 Polarographic
Electrodes
BB-34090820
BB-34090821
BB-34090822
BB-34090823
BB-34090824
BB-8848661
BB-8848662
BB-8848663
BB-8848664
BB-8848665
pO2 Electrode 12/120-L-HM UniVessel® 0.5 L, for BIOSTAT® Qplus
Polarographic electrode (Hamilton)
y Electrode length 120 mm
pO2 Electrode 12/160-L-HM UniVessel® 1 L, for BIOSTAT® Aplus, Bplus,
Qplus
Polarographic electrode (Hamilton)
y Electrode length 160 mm
pO2 Electrode 12/215-L-HM UniVessel® 2 L, for BIOSTAT® Aplus, Bplus
Polarographic electrode (Hamilton)
y Electrode length 215 mm
pO2 Electrode 12/325-L-HM UniVessel® 5/10 L (5 L min. AV),
for BIOSTAT® Aplus, Bplus
Polarographic electrode (Hamilton)
y Electrode length 325 mm
pO2 Electrode 12/425/2-L-HM UniVessel® 10 L (1.5 L min. AV),
for BIOSTAT®, Bplus
Polarographic electrode (Hamilton)
y Electrode length 425 mm
pO2 Electrode 12/120-VP-HM UniVessel® 0.5 L, BIOSTAT® B-DCU II
y Polarographic electrode (Hamilton)
y Electrode length 160 mm
pO2 Electrode 12/160-VP-HM UniVessel® 1 L, BIOSTAT® B, B-DCU II
y Polarographic electrode (Hamilton)
y Electrode length 160 mm
pO2 Electrode 12/225-VP-HM UniVessel® 2 L, BIOSTAT® B, B-DCU II
y Polarographic electrode (Hamilton)
y Electrode length 225 mm
pO2 Electrode 12/325-VP-HM UniVessel® 5 L, 10 L (5 L min. AV),
BIOSTAT® B, B-DCU II
y Polarographic electrode (Hamilton)
y Electrode length 325 mm
pO2 Electrode 12/425-VP-HM UniVessel® 10 L (1.5 L min. AV),
BIOSTAT® B, B-DCU II
y Polarographic electrode (Hamilton)
y Electrode length 425 mm
pO2 electrodes from Mettler-Toledo are available on request!
Electrodes and Sensors
59
Connecting cable for individual sensors, probes and electrodes:
(The cables are included in the above-mentioned probe sets for Aplus, Bplus, Qplus
and B-DCU I)
4.4.1.2 Optical pO2 Electrodes
BB-8848680
VP8|VP8 for polarographic pO2 sensor BIOSTAT® B, B-DCU II
BB-8848668
pO2 electrode - 12/120/VP UniVessel® 0.5L BIOSTAT® B-DCU II
y Optical electrode (Hamilton)
y Electrode length 120 mm
pO2 electrode - 12/160/VP UniVessel® 1L
BIOSTAT® B, B-DCU II
y same as BB-8848668 with electrode length 160 mm
pO2 electrode - 12/225/VP UniVessel® 2L
BIOSTAT® B, B-DCU II
y same as BB-8848668 with electrode length 225 mm
pO2 electrode - 12/325/VP UniVessel® 5L / 10L (5L min. AV)
BIOSTAT® B, B-DCU II
y same as BB-8848668 with electrode length 325 mm
pO2 electrode - 12/425/VP UniVessel® 10L (1,5L min. AV)
BIOSTAT® B, B-DCU II
y same as BB-8848668 with electrode length 425 mm
BB-8848669
BB-8848670
BB-8848671
BB-8848672
Connecting cable for individual sensors, probes and electrodes:
BB-8848685
VP8|VP8 for optical pO2 sensor BIOSTAT® B, B-DCU II
4.4.2 Operating Information
4.4.2.1 Assembly and Connection
1. Insert the pO2 electrode into the appropriate lid port t “Vessel drawing,
recommended arrangement of attachment parts” and hand tighten firmly.
When gassing the culture vessel through a mirco sparger t Section “Micro sparger”,
align it or use a cover port to ensure that the electrode is not located directly above
the bubble flow. Directing the flow of input gas right at the membrane can disrupt the
pO2 measurement.
O-ring
Installing the pO2 electrode
60
Electrodes and Sensors
2. Remove the connecting cable for sterilization, it may not be autoclaved.
3. To protect the plug against the impact of steam in the autoclave, you cover it with
the protective cap included in the delivery or wrap some aluminum foil around.
4. After autoclaving and setting up the culture vessel at the workplace and|or before
starting the process, connect the electrode on the measurement and control
system. When using bioreactors with multiple culture vessels (BIOSTAT® Bplus
Twin, Qplus, B-DCU), note which connection socket is assigned to the respective
culture vessel.
5. Calibrate the pO2 electrode, as described below and in the t “Documentation on
the measurement and control system”.
6. Configure the pO2 measurement and control parameters as described in the
t “Documentation on the measurement and control system”.
4.4.2.2 Calibrating
Polarographic Electrodes
Prior to calibration, the pO2 electrode must be polarized for at least 2 h.
You must repeat the polarization if the electrode has been disconnected from the
amplifier for about 10 minutes, but this doesn’t take that long t “Manufacturer’s
documentation”.
For polarization, you can use a polarization module from the manufacturer or connect
the pO2 electrode on the basic unit and|or measurement and control system and then
switch it on at the beginning of the calibration.
After sterilization in the autoclave, calibrate the pO2 electrode in the culture vessel.
First determine the electrode´s current when no oxygen is present in the medium: this
is the “zero point”. Then, determine the current after saturation with oxygen as the
“slope”. You will find instructions on what entries to make in the operating menus in
the t Documentation on the measurement and control system.
1. You can determine the zero point before you gas the culture vessel with air and|or
oxygenated gas. Heating during sterilization will normally degass the culture
medium to such an extent that the any residual oxygen will still be within the
accuracy range of the electrode.
If this is no (longer) possible, you can gass the culture medium with nitrogen
(N2 with 99.98% purity), until the dissolved oxygen is completely displaced
(constant reading display around 0% pO2).
2. To the calibrate the slope and configure the measuring range, connect the air
and|or gas supply and gass the culture medium. Adjust the gas supply as intended,
like as a reference value for 100% pO2. t Documentation on the measurement
and control system or on the gassing module used.
3. After calibration, you can adjust the gas supply as intended for process start.
4.4.2.3 Calibrating
Optical Electrodes
The long polarization time is not necessary when using optical electrodes.
After autoclaving, calibration can be started immediately. VISIFERM DO sensors are
equipped with a VP8 plug head. The eight golden contacts are designated Pin A, Pin B,
... and Pin H. For easy assignment of the pins, the plug head has been coded between
Pin A and Pin B. During autoclaving, this VP connection must also be protected by a
cap or other heat-resistant cover.
After autoclaving, connect the sensor and the bioreactor with the connecting cable.
The easiest and safest way to connect the VISIFERM DO sensors is using HAMILTON VP8
cables, which are part of the equipment supplied with the BIOSTAT® bioreactor.
The calibration process is the same as for the polarografic electrodes.
Further information on the calibration routine can be found in the HAMILTON
manufacturer's documentation and in the operating manual for your BIOSTAT®
bioreactor.
Electrodes and Sensors
61
4.5 Redox Electrode
Sterilizable combination electrodes are available for the measuring the Redox value.
4.5.1 Equipment and
Specifications
BB-8841222
BB-8844196
BB-8844186
BB-8844194
Redox electrode 1 L, complete, for BIOSTAT®Qplus
y Pressurized pasteous electrode for installation in 12 mm lid port
y Electrode length 120 mm
y Connection cable 100 cm
y Bottle of Redox buffer, 250 ml
Redox electrode 1 L, complete, for BIOSTAT®Qplus
y Electrode length 160 mm
Redox electrode 2 L, complete, for BIOSTAT®Qplus
y Electrode length 200 mm
Redox electrode 5/10 L (5 L min. AV), complete, for BIOSTAT®Bplus
y Electrode length 325 mm
On the B and B-DCU II, combination electrodes are used during Redox measurement
for the simultaneous determination of pH and Redox potential:
BB-8844200
pH & Redox electrode, 120/12-VP electrode 0.5 L, BIOSTAT® B-DCU II
y Pressurized pasteous electrode for installation in 12 mm lid port
y Electrode length 120 mm
BB-8844201
pH & Redox electrode, 160/12-VP electrode 1 L, BIOSTAT® B-DCU II
y Pressurized pasteous electrode for installation in 12 mm lid port
y Electrode length 160 mm
BB-8844202
pH & Redox electrode, 225/12-VP electrode 2 L, BIOSTAT® B-DCU II
y Pressurized pasteous electrode for installation in 12 mm lid port
y Electrode length 225 mm
BB-8844203
pH & Redox electrode, 325/12-VP electrode 5 L / 10 L (5 L min. AV),
BIOSTAT® B-DCU II
y Pressurized pasteous electrode for installation in 12 mm lid port
y Electrode length 325 mm
BB-8844204
pH & Redox electrode, 325/12-VP electrode 1.5 L / 10 L (5 L min. AV),
BIOSTAT® B-DCU II
y Pressurized pasteous electrode for installation in 12 mm lid port
y Electrode length 425 mm
Connecting cable for individual sensors, probes and electrodes:
(The cables are included in the above-mentioned probe sets for Aplus, Bplus, Qplus
and B-DCU I)
BB-8848681
VP8|VP8 for pH&Redox sensor BIOSTAT® B, B-DCU II
4.5.2 Operating Information
4.5.2.1 Reactivating
Redox Electrodes
1. Before first use or after it has dried out after long-term storage or after dry
storage, you must carefully reactivate your Redox electrode.
Glass membranes, as used in pH electrodes, form a thin aqueous gel layer that
determines the measuring properties (response time, slope, alkaline error etc.).
This layer needs to regenerate.
2. Follow the instructions in the t “Manufacturer’s documentation”. You can place
the Redox electrode into a beaker with demineralized water for about 12 ... 14 h.
4.5.2.2 Calibrating the
Redox Electrode
The Redox electrode does not need to be calibrated. It can be checked with the
standard buffer supplied by the manufacturer. The Redox value measured must
correspond with the reference value on the buffer bottle.
Check the Redox electrode prior to installation in the culture vessel.
62
Electrodes and Sensors
4.5.2.3 Assembly and
Connection
Danger of damaging the pH electrode.
Other attachment parts in the culture vessel, particularly the rotating stirrers,
may not butt up against the electrode. Use a suitable lid port, e.g. one located
further away.
1. Insert the Redox electrode into a suitable free lid port.
The insertion depth depends on the culture vessel and shaft length. For 10 L vessels,
there are electrodes with different lengths, depending on the minimum operating
volumes.
2. Screw in the Redox electrode fitting hand tight. Never use a pliers or wrench!
3. To prevent the pH electrode from drying out, fill the culture vessel with water or
culture medium after you have installed all parts.
4. The cable must not be autoclaved. To protect the sensor plug from the effects of
steam during autoclaving, you can attach the protective cap supplied or wrap
aluminum foil around the plug.
5. After autoclaving and setting up the culture vessel at the workplace and|or before
starting the process, connect the electrode on the measurement and control
system.
When using bioreactors with multiple culture vessels (BIOSTAT® Bplus Twin, B Twin,
Qplus, B–DCU), note which measurement amplifier panel is assigned to the respective
culture vessel.
Redox Electrode
4.6 Turbidity Electrode
FUNDALUX® II
FUNDALUX® turbidity measurement systems to determine optical density can be
purchased optionally. Further details can be found in the t “Operating manual
Fundalux®”.
External Accessories
63
5. External Accessories for
Installation and Connection
5.1 Exhaust Cooler
5. External Accessories for Installation
and Connection
Exhaust coolers reduce the moisture content of the exhaust. The cooler features a
jacket for circulating the cooling water. The exhaust moisture condenses on the inner
tube that the cooling water flows around. This reduces the risk that the exhaust filter
gets blocked and that moisture is loss through evaporation, particularly during longterm processes.
If the exhaust filter is blocked, the culture medium will be gassed insufficiently.
This can lead to an inadmissible rise in the internal pressure inside the culture
vessel can or to its unsterile release through connections of the lid installations
(if they have been installed improperly or if the seals are defective).
If the vessel is assembled with an optional extension kit, you can reposition the
exhaust cooler when you place the culture vessel in the autoclave. In this case,
an autoclave with a correspondingly lower internal compartment can be used to
accommodate the culture vessel.
5.1.1 Equipment and
Specifications
BB-8841208
BB-8846871
BB-8846880
BB-8846905
BB-8846906
BB-8846907
Exhaust cooler for UniVessel® 0.5 L, for BIOSTAT® Qplus
y Connection PG 13.6 d 4
y Install in d 12 mm lid port
Exhaust cooler for UniVessel® 1/2 L, for BIOSTAT® Aplus, Qplus
y Cooling water connection: Fabric-reinforced silicone tubing
with quick connect couplings
y Install in d 19 mm lid port
Exhaust cooler for UniVessel® 5 L, for BIOSTAT® Aplus
y Install in d 19 mm lid port
y Equipment, installation in cover plate as above
Exhaust cooler for UniVessel® 0.5 L, for BIOSTAT® B-DCU II
y Install in d 12 mm lid port
Cooling fingers UniVessel® 1/2 L, for BIOSTAT® B, Bplus, B-DCU II
y Install in d 19 mm lid port
Exhaust cooler for UniVessel® 5/10 L, for BIOSTAT® B, Bplus, B-DCU II
y Install in d 19 mm lid port
An extension kit is optionally available for height adjustment of the exhaust
cooler in the autoclave (see also t “Section 5.1.2.2")
For height adjustment dimensions, please refer to Table t “2.3.2”.
64
External Accessories
5.1.2 Operating Information
5.1.2.1 Assembly on the Culture
Vessel without Extension Kit
1. Insert the exhaust cooler in the lid port and screw it tight.
2. Mount the sterile exhaust filter via a piece of silicone hose t “Assembly of the
exhaust filter”.
3. After sterilization in the autoclave, connect the cooling water supply and exhaust.
Note the markings for the inlet and return to the tubing adapters.
O-ring (2)
(1) O-ring (2) head piece exhaust cooler B5, 18.77 +1.78, EPDM | FDA
(2) O-Ring (1) lid port fitting, 15.6 +1.78, EP
O-ring (1)
Exhaust cooler for 2 L culture vessel
(figure shown as an example for the
BIOSTAT® Aplus version)
External Accessories
65
5.1.2.2 Installation with
Extension Kit
1. Insert the tubing adapter (2) of the extension kit (1) in the lid port intended for
the exhaust cooler and screw it tight.
2. Replace the nearest screw connection on the lid using the screw connection (4)
with the support rod pin for the extension kit.
3. Insert the support rod (3) pin and fasten it with the fastening screw (5).
4. Insert the upper tubing adapter (6) of extension kit in the hole on the support rod
and fasten it with the fastening screw.
5. Screw the exhaust cooler (7) into the adapter at the top of the piece of hose.
6. Connect the exhaust filter via a piece of silicone hose to the exhaust cooler
t “Assembly of the exhaust filter”.
7. After sterilization of the culture vessel, connect the cooling water supply and
drain before you start the process.
O-ring
Installation with Extension Kit
5.1.2.3 Folding up the
Exhaust Cooler
1. Remove the support screw holding the exhaust cooler adapter (6).
2. Remove the adapter from the support.
3. Turn the exhaust cooler over.
4. After autoclaving and setting up the culture vessel in the work area, insert the
adapter with the exhaust cooler in the support holes again and fix it in place with
the fastening screw.
5.1.2.4 Connecting the
Cooling Water Supply
1. Connect the tubing for the cooling water supply and return from the basic unit |
temperature control module to the exhaust cooler. Observe the markings for
intake and return.
2. Open the cooling water supply when activating the culture vessel gassing.
3. Cooling water is fed through the exhaust cooler at a constant flow rate,
as predetermined by the valves in the basic unit of the bioreactor or on the
temperature control module | connection to the laboratory cooling water supply,
according to the documentation on t “Installation and Operation of the Basic
Unit | Temperature Control Module”.
It is recommendable to use a cooling thermostat, e.g. a continuous flow cooler,
in the case of high cooling water consumption or long-term processes.
66
External Accessories
5.2 Sterile Filters for Air |
Gas Supply and Exhaust
The inlet or exhaust filters are autoclavable membrane filters in plastic housings.
The pore size of approx. 0.2 μm ensures the sterile supply of air | gas to the culture
vessel and sterile ventilation.
y The filter is necessary for the following connections:
– Gas supply by way of the sparger pipe in the culture vessel (sparger)
– Gas supply through silicone hose membrane when equipped with aeration
basket for bubble-free aeration t “Membrane aeration system"
– Gas supply to the “headspace”, e.g. in the “exclusive flow” model of the
bioreactor gassing module
– Exhaust from the headspace
– Exhaust from silicone hose membrane when equipped with aeration basket
17805—————E
5.2.1 Equipment and
Specifications
BB-8846928
Inlet | exhaust filter Midisart 2000 (12 pcs.)
Disc filter for use as inlet | exhaust filter
y PTFE filter in polypropylene housing
y Pore size 0.2 μm
y Filter area 20 cm2
y Flow rate at 0.1 barg 5 L/min
y Autoclavable at 121°C min. 20+
y Hose nozzles for hoses on both sides starting at d ID = 3.2 mm
Exhaust filter Sartofluor capsule
Filter capsule for use as exhaust filter
y PTFE filter in PP capsule
y Pore size 0.2 μm
y Filter area 0.03 m2
y Hose nozzles for hoses on both sides starting at d ID = 4 mm
5.2.2 Operating Information
5.2.2.1 Assembling and
Connecting the Inlet Air Filter
Pay attention to the flow direction of the respective gas mixtures when
assembling the filter.
Pay attention to the “IN” markings on the filters!
They mark the gas input side.
If you do not clamp off the hose (2), culture medium can be forced into the
tubing by way of the sparger pipe during sterilization and block the filter.
1
1. Connect the air inlet filter (2) with a piece of silicone hose to the sparger pipe in
the culture vessel (or to the inlet to the membrane aeration basket, if installed).
2. Before sterilization in the autoclave, clamp off the hose with a hose clamp.
3. If headspace aeration is intended, connect a second filter to a universal adapter
extending into the headspace. It is not necessary to clamp the hose.
2
4. Use a sufficiently long piece of hose to connect the basic unit or gassing module
of the bioreactor to the hose nozzle (1).
Membrane sterile filter
External Accessories
67
5.2.2.2 Assembling and
Connecting the Exhaust Filter
1. Connect a short section of silicone hose (3.2+1.6 mm) to the hose nozzle (2).
2. Connect the free end of the silicone hose to the hose nozzle on the exhaust cooler.
3. It’s not necessary to connect a hose to the open outlet (1). The exhaust can blow
freely into the room. Exception: You wish to connect an exhaust analysis.
Do not clamp off exhaust air path hose. The exhaust air path serves to
compensate for ambient pressure during heating and cooling.
Do not allow hose to come loose unintentionally. If unsterile air gets into the
culture vessel, ambient germs can disrupt the process.
Check the exhaust air path after sterilization. Foaming medium may have
entered into the exhaust air path. The filter may have to be replaced.
5.2.2.3 Exhaust Filter in the
Membrane Aeration Basket
1. Connect a piece of silicone hose to the hose nozzle (2) on the universal adapter
in the cover plate, which is connected to the outlet of the membrane aeration
basket.
2. Use the exhaust cooler and connected filters for deaeration of the headspace.
3. After sterilization and setup at the work place, fasten the pressure hold unit
t “Membrane aeration system to the culture vessel” and connect the exhaust
filter (1) to it.
5.2.2.4 Connecting the Bioreactor
Gas is supplied to the culture vessel by way of the control unit or the gassing module.
1. Connect the air inlet filter with the sparger outlet to the control unit | gassing
module.
2. Connect the headspace filter to the outlet “overlay” for headspace aeration
(only in versions with the gassing module “exclusive flow").
68
External Accessories
5.3 Cooling Fingers
Cooling fingers serve to cool single walled culture vessels.
The cooling water supply is regulated using the temperature control on the bioreactor.
The cooling fingers can also be connected to separate cooling devices in the case that
cooling water alone does not provide sufficient cooling.
5.3.1 Equipment and
Specifications
BB-8846456
BB-8847819
BB-8847827
BB-887818
BB-8847822
BB-8847823
BB-8847824
Cooling fingers for UniVessel® 1 L, for BIOSTAT® Aplus, Qplus
y Installation in d 19 mm lid port:
– Insertion depth HE = 160 [mm], dip tube d T = 12 mm
y Connection via quick connect couplings and fabric-reinforced
silicone hoses
Cooling fingers for UniVessel® 2 L, for BIOSTAT® Aplus
y Like cooling fingers UniVessel® 1 L, insertion depth HE 210 mm,
dip tube d T = 18 mm
Cooling fingers for UniVessel® 5 L, for BIOSTAT® Aplus
y Like cooling fingers UniVessel® 1 L, insertion depth HE 310 mm,
dip tube d T = 18 mm
Cooling fingers for UniVessel® 1 L, for BIOSTAT® Bplus, B, B-DCU II
y Install in d 19 mm lid port:
– Insertion depth HE = 160 [mm], dip tube d T = 12 mm
y Connection via quick connect couplings and fabric-reinforced
silicone hoses
Cooling fingers for UniVessel® 2 L, for BIOSTAT® Bplus, B, B-DCU II
y Like cooling fingers UniVessel® 1 L, insertion depth HE 210 mm,
dip tube d T = 18 mm
Cooling fingers for UniVessel® 5 L, for BIOSTAT® Bplus, B, B-DCU II
y Like cooling fingers UniVessel® 1 L, insertion depth HE 310 mm,
dip tube d T = 18 mm
Cooling fingers for UniVessel® 10 L, for BIOSTAT® Bplus, B, B-DCU II
y Like cooling fingers UniVessel® 1 L, insertion depth HE 425 mm,
dip tube d T = 18 mm
External Accessories
69
5.3.2 Operating Information
The hoses (2) for connecting the bioreactor or a separate cooling device are attached
to the cooling finger via quick connect couplings. The adapter on the hoses are
labeled:
1
2
3
y Supply: Hose with quick connect coupling (3)
y Return: Hose with plug-in sleeve (1).
Hoses for connecting the basic unit are supplied as described in the t “Temperature
control module for single walled vessels”. If you wish to connect other cooling devices,
be sure to use hoses of sufficient length.
HE
Replacing Defective Hoses
1. Install the plug-in sleeve on the return and the coupling on the supply hose.
Secure the hoses against slipping.
2. Fasten the hose with the coupling to the supply of the cooling finger and the hose
with the plug-in sleeve at the return. Observe the markings.
DT
Cooling fingers, example UniVessel® 2 L
for BIOSTAT® Aplus
5.3.2.1 Assembly on the Culture Vessel
1. Screw the cooling finger into a free 19 mm port d in the cover plate of the
culture vessel. Carefully hand tighten the cap nut.
2. Fasten the hoses to the culture vessel before transporting the culture vessel and
placing it in the autoclave.
3. Check that the cooling finger is firmly seated in the cover plate after autoclaving.
5.3.2.2 Connecting and Operating the Cooling Water Supply
Operating pressure of separate cooling cycle for cooling fingers only:
max. 1.5 barg.
The cooling water supply via a separate cooling cycle must be regulated
manually. In that case, the temperature regulation of the bioreactor only
affects heating.
Condensation may form at very low temperatures.
Make sure that condensation water cannot cause disruptions or damage
equipment.
After autoclaving and setting up the culture vessel at the work place:
1. Connect the cooling water hoses to the control unit of the bioreactor (or to
the temperature control module for single walled culture vessels). Observe the
markings for supply and return.
2. When directly connecting the hoses to external cooling devices or cooling
circulation systems, use the proper adapters. The connections must not be
allowed to loosen unintentionally under pressure.
3. Set the pre-pressure of the cooling water supply t “P&I Diagram”.
70
External Accessories
5.4 STT Quick Connector
The STT quick connect coupling can be used to produce fast and secure 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
culture vessel line, the male connector to the supply vessel transfer line or the harvest
container.
5.4.1 Equipment and
Specifications
BB-8809240
BB-8809208
BB-8809410
BB-8809402
STT quick connect coupling half (female)
y Autoclavable quick connect coupling half made of stainless steel
y for sterile connection of hoses with an internal diameter of
3.2 ... 5.0 mm
y Equipment supplied: Female coupling half, 1 dummy plug
STT quick connect coupling half (male)
y for sterile connection of hoses with an internal diameter of
3.2 ... 5.0 mm
y Equipment supplied: Quick connect coupling, male half, 1 cap
STT quick connect coupling half (female)
y for sterile connection of hoses with an internal diameter of
1.6 ... 2.0 mm
STT quick connect coupling half (male)
y for sterile connection of hoses with an internal diameter of
1.6 ... 2.0 mm
External Accessories
71
5.4.2 Operating Information
5.4.2.1 Assembly
Prepare the reservoir vessel and the connection to the culture vessel in such a way
that the STT plug-in nipple is positioned at the line to the media vessel and the STT
coupling at the connection to the culture vessel. Connect the STT components prior
to sterilization.
2
1
3
Assembly and Connection of the Exhaust Filter (7):
1. Place the slit membrane (2) into the membrane holder (1).
6
4
7
2. Screw the adapter (3) onto the membrane holder (1).
5
3. Cut a piece of silicone hose (4) and attach it to the coupling part and to the
addition ports on the culture vessel.
Prepare and connect coupling part
of the STT coupling
4. Insert the dummy plug (5). This maintains sterility after autoclaving until the plug
is connected.
5. Autoclave STT coupling and culture vessel.
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.
6
9
10
8
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.
Connect the male part of the STT coupling
5.4.2.2 Connecting the STT
Quick Connector
1. Remove dummy plugs from the 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.
6
7
8
Connect STT coupling parts
5.4.2.3 Transferring a Medium
1. To transfer the media, lift reservoir vessel or connect the hose to a pump.
2. A hose pump can be used to extract the medium and transfer a sample 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.
72
External Accessories
5.5 Bypass Sampler
The bypass sampler allows you to remove culture medium from the culture vessel, then
route it through a membrane holder and back into the vessel, all within in a closed and
sterile system. The membrane holder contains a self-closing inoculation membrane
which can be pierced, e.g. with an injection syringe.
In this way, the bypass sampler can be used to:
1. Remove small sample volumes, e.g. for off-line analysis of media components for
which you require the most current and representative samples possible.
2. Introduce media whose effect on the culture you would like to examine, e.g.
vitamins, serums or special inductors | inhibitors of the metabolic cell functions.
5.5.1 Equipment and
Specifications
BB-8844348
5.5.2 Operating Information
Assembly and Connection
1. Cut the teflon tube or a piece of silicone hose, as sampling tubing, to a sufficient
length that allows you to take representative samples of the medium
compartment at a suitable height in the culture vessel.
Membrane holder
Return
Bypass sampler M | B
y Membrane holder with inoculation membrane
y 2-channel screw socket 19 mm for assembly on the culture
vessel
y 2 m silicone hose 1.6 +1.6 mm for the bypass line
y 0.25 m PTFE hose 3.5 +1.0 mm as sample rising pipe
Supply
Peristaltic pump
Instead of a 2-channel screw socket, you can connect the dip tube via a free
adapter or inoculation port and route the medium back into the culture vessel.
Harvesting pipe
2. Ensure that external hoses connecting the membrane holder and the return are of
sufficient length to use a hose pump for transferring the culture medium through
the bypass.
Culture vessel
Setting up the bypass
Return
Sampling
O-ring
2-channel
adapter
Cover port
d 19 mm
Preparing the sampling equipment
1. When using a 2-channel screw socket: Insert the sampling tubing (the pipe) used
to remove media from the culture vessel onto one of the hose nozzles from below.
Screw the nozzle into one 19 mm lid port.
2. Mount the hose to the membrane holder on the hose nozzle above the sampling
tubing of the 2-channel screw socket | dip tube of the culture vessel.
3. The return hose from the membrane holder can be connected to the hose nozzle
of the 2-channel screw socket, to a free lid port or to an inoculation nozzle.
Teflon pipe
or silicone hose
2-channel screw socket
External Accessories
73
Cap
O-ring
Cap nut
Preparing the Membrane Holder
1. Place the slit membrane in the basic adapter and fasten it with the disc and the
cap nut.
2. Put the protective cap on.
Slit membrane
3. Connect the bypass line. Before sterilization in the autoclave, clamp off the hose
with a hose clamp.
Basic adapter
4. The membrane holder and bypass can be autoclaved with the culture vessel.
Disc
Silicone hose
Membrane holder of the bypass sampler
Filling with Media | Sampling
1. Prepare a syringe of sterile substance for adding media. Use an empty or sterile
syringe for sampling.
Cap
Injection syringe
Disc
Cap nut
Slit membrane
2. Pump the culture medium through the bypass for a while to get a sample.
This way you can obtain a representative sample.
3. Remove the cap from the bypass sampler. Flame the membrane or moisten it with
disinfecting agent.
Basic adapter
4. Pierce the membrane with the syringe and inject the medium or extract the
sample. Remove the syringe and place the cap back on.
Piercing the bypass sampler
Flaming the inoculation membrane should be preferred over the use of
disinfecting agents. Disinfecting agents, e.g. alcohols, may contain spores
which can contaminate the culture or residue may enter the bypass when
piercing the membrane.
5.6 Other Accessories
74
External Accessories
When assembling other accessories, e.g.
– Heating jackets and
– Drive motors
proceed according to the t “Installation Instructions for Bioreactor Control"
6. Equipping the Vessel
Before Process Start
6.1 Corrective Solution Bottles
6. Equipping the Vessel Before
Process Start
The are a variety of storage bottles (also “corrective solution bottles") available for
different corrective solutions or substrates.
When large volumes are needed, you can use reservoir | harvesting bottles
(balloon bottles made of polypropylene) with a volume of 10 L, 20 L or 50 L.
6.1.1 Equipment and
Specifications
In general, all corrective solution bottles feature
– a headpiece made of stainless steel AISI 316 L (1.4435)
and a
– ventilation filter 0.2 μm.
Autoclavable glass bottle made of borosilicate glass,
with two hose nozzles for 3.2 +1.6 mm hose
y Screw cap with silicone gasket
y PTFE hose, acid-, lye- and temperature-resistant
BB-8823600
Reservoir bottle 250 mL
BB-8841234
Reservoir bottle 250 mL 4 ports
BB-8823618
Reservoir bottle 500 mL
BB-8823675
Reservoir bottle 1000 mL
Corrective solution bottle 250 mL
6.1.2 Operating Information
Autoclavable glass bottle made of polypropylene,
with 3 hose nozzles for 3.2 +1.6 mm hose
y Lid screw connections with silicone seal
y Silicone hose 3.2 + 1.6 mm, 40 cm length
BB-8823642
Supply | harvesting bottle 10 L
BB-8823650
Supply | harvesting bottle 20 L
BB-8823669
Supply | harvesting bottle 50 L
Danger of injury from shattered glass!
Danger of chemical burns from acids and lyes!
Damaged glass bottles can shatter during autoclaving or handling and media
e.g. acids or lyes, may be released unintentionally. Media may also be released
unintentionally due to damaged hoses.
Handle glass bottles with care. Replace damaged bottles.
Regularly check silicone seals and hoses and the ventilation filters for damage,
and replace them regularly.
Protect yourself against caustic burns. Wear safety gloves and safety glasses.
All equipment must be resistant to the corrective solutions used. If this has not
been guaranteed, their resistance must be checked.
Avoid the use of hydrochloric acid (HCl) for adjusting pH. HCl can also attack
stainless steel parts.
Make sure the total volume of the culture vessel cannot be exceeded by the
additional volume of the storage bottles. You may connect an empty harvest
container to a dip tube for safety reasons, so that potentially overflowing media
can safely flow out.
Vessel Equipment
75
6.1.2.1 Preparing and
Connecting the Bottles
A number of bottles should be prepared for longer or continuous processes in order to
have sufficient sterile solution available.
1. Insert the Teflon tube (7) onto a hose nozzle. Shorten it until it reaches to about
1-2 mm above the floor of the bottle.
5
6
4
2. Fill the bottle (1) with anti-foam solution, acid, alkaline, or substrate. Place the
silicone gasket (2) and the headpiece (3) onto the edge of the glass and close the
bottle with the screw cap (4).
3. Place a piece of silicone hose (6) onto the hose nozzle to which the Teflon pipe (7)
is attached.
3
7
2
The transfer hose must be long enough that you can insert it into the associated
peristaltic pump.
4. Place the sterile filter (5) and silicone hose onto the remaining hose nozzle on the
bottle.
5. Secure all hoses with hose clamps.
Connecting the Bottles
1. Connect the transfer hoses to the inlet in the cover plate t “Quadruple Addition
Ports” or t “Universal Adapter”. Secure all hose connections with hose clamps.
1
2. If you have not yet done so, mount the bottle holder to the culture vessel and
insert the bottles t “Bottle Holder”.
3. Before sterilization, clamp off the hose with hose clamps.
In the case of overpressurization in the bottles, no media may be forced into the
hose | culture vessel.
4. Autoclave the corrective solution bottles together with the culture vessel.
5. Set up the culture vessel with the corrective solution bottles at the work place.
Connecting Additional Corrective Solution Bottles In-Process
Carefully ensure sterility when connecting additional, separately autoclaved bottles to
the culture vessel in order to avoid contaminating the culture. You can connect these
bottles easily and securely via a t “STT quick coupling”.
Installation in the Peristaltic Pumps
76
Vessel Equipment
6.2 Manual Sampler
The manual sampler allows discontinuous sampling from the culture vessel.
6.2.1 Equipment and
Specifications
The manual sampler, order no. BB-8844623, contains a sampling tube, volume 15 mL
(2), a suctioning syringe with 0.2 μm, filter d 25 mm as injection attachment (4) and
the clamp holder to fasten the support rod of the culture vessel.
Additionally, there are silicone hoses (sampling tubing d 3.2 +1.6 mm, a hose for
syringe d 1.6 +1.6 mm) and a hose clamp. The cover of the collection tube has 3 hose
nozzles for the sampling tubing from the culture vessel (1), for the sampling outlet (5),
and for the syringe (3). The sterile filter on the syringe prevents unsterile air from
entering while suctioning and injecting samples.
Vessel Equipment
77
6.2.2 Assembly Information
1. Unscrew the cap on the sampling tube. Insert a bit of silicone hose d 3,2 +1.6 mm
onto the bottom of the rising pipe that reaches the bottom of the tube. In this
way, the tube can be fully emptied when injecting a sample. Finally, screw the cap
back on.
2. Insert a piece of silicone hose d 3.2 +1.6 mm (5) onto the hose nozzle with the
rising pipe. This serves to eject the sample from the collection tube.
3. Insert a piece of silicone hose d 3.2 +1.6 mm (1) onto the hose nozzle of the
culture vessel and on the hose nozzle of the sampling rising pipe for the
connection to the culture vessel. This line serves for suctioning off the sample.
4. Insert the sterile filter (7) onto the syringe and connect it to the unoccupied hose
nozzle on the sampling tube with silicone hose d 1.6 +1.6 mm (3).
5. Clamp the holder for the hand sampler to a support rod (11) on the culture vessel
with a fastening screw (19). Insert the sampling tube and the syringe into the
holder.
6. Clamp off the line to the culture vessel (1) with a hose clamp before autoclaving,
so that no medium can be forced in the sampling tube from overpressurization in
the culture vessel. Also clamp off the drain line (5).
7. Autoclave the completely assembled hand sampler along with the culture vessel.
Hand sampler:
- above: side view
- below: view from above
78
Vessel Equipment
6.2.3 Operating Notes
for Sampling
1. Press the plunger of the syringe to prepare a sample extraction using an open
outlet hose (5).
2. Remove the hose clamp from the hose connecting the culture vessel and the
handle sampler (1). Use this to clamp off the outlet hose (5).
3. Pull out the syringe plunger (4). This way you can suction the sample out of the
culture vessel via the rising pipe. Transfer as much sample material as is needed
into the sampling tube (2). Then press the syringe plunger back in a bit, so that the
sampling material remaining in the connection hose finds its way back into the
culture vessel.
4. Take the hose clamp off the outlet tubing and clamp off the connecting hose to
the culture vessel (1).
5. Slowly press in the syringe plunger to transfer the sample into an external holding
tank for further processing. The sample material is pressed into the outlet tubing
through the rising pipe in the collection tube (5).
As described in step 5, empty the outlet tubing as much as possible. Residue in
the tube may contaminate samples taken later and adversely influence
subsequent analyses.
The sterile filter injection attachment prevents unsterile air from getting into the
sampling tube via the syringe
6.3 Bottle Holder
The bottle holder is simply mounted to the stainless steel frame so that the storage
bottles can be inserted.
BB-8841232
Culture vessel tray for 3+0.5 L UniVessels®
BB-8846464
Bottle holder for UniVessel® 1 L
BB-8847428
Bottle holder for UniVessel® 2 L
BB-8847436
Bottle holder for UniVessel® 5 L
BB-8847444
Bottle holder for UniVessel® 10 L
Bottle holder
Bottle holder Qplus
Vessel Equipment
79
For your notes
80
Notes
7. Appendix
7. Appendix
7.1 Cleaning and Maintenance
The frequency and method of cleaning and maintenance depend on the degree to
which the components in the culture medium, cells and metabolic products, such as
proteins, adhere to the vessels and their internal equipment.
7.1.1 Measures after Previous Use
Possible biohazards, depending on nature of the cultivated microorganisms and
cells.
Observe the safety regulations relating to your process on the handling of
microorganisms or cells and the devices contaminated by them.
After harvesting product, you can fill the culture vessel with water and autoclave
it once more.
If no special safety regulations apply, you can rinse the culture vessel with water.
In the case of short breaks until the next process is started, you can fill the vessel with
water to prevent the electrodes from drying out.
The culture vessel needs to be dismantled and under basic cleaning only when
retrofitting the equipment, for maintenance as well as for longer breaks in operation.
Disassembly may be necessary if you want to cultivate different microorganisms and |
or cells in a subsequent process.
Replace the glass vessels as well as O-rings and seals on the equipment if they
show damage or stubborn fouling that cannot be removed with the following
measures:
Cleaning glass vessels after disassembly in a conventional laboratory dishwasher.
Removing cell deposits and cell debris on the glass with laboratory glass cleaning
agents or mechanically. Removing inorganic deposits using diluted hydrochloric
acid or similar cleaning agents.
Clean metal parts, gaskets, O-rings mechanically, using alcohols or with mild
(non-abrasive or corrosive) cleaning agents. When using chemical agents, you
must rinse all parts thoroughly with water after cleaning.
For the cleaning and maintenance of sensors, follow the t“Manufacturer’s
instructions”.
7.1.2 Culture Vessel
Disassembly
The O-ring can stick to the vessel lid. Take care not to drop or bump the glass
vessel and|or damage any of the built-in sensors and accessory parts.
In the case of double walled culture vessels, you can remove the glass vessel from its
stand as follows:
1. Remove the hose adapters connected to the temperature control circulation from
the glass ports on the double wall.
2. Detach and remove all equipment and attachment parts accessible from the
outside (motor, sensors, signal cables, tubing connections, exhaust cooler, etc.).
Unscrew the screw connections on the lids. Turn the lid slightly and then lift it up
carefully from the glass vessel.
3. Lift up the glass vessel with the stand carefully and turn it upside-down with the
glass flange and | or stand ring onto a clean surface, e.g. a rubber mat.
4. On the stand ring, detach the screws to the attachments holding the glass vessel.
Lift the stand up and out over the glass vessel without bumping the glass ports.
Appendix
81
In the case of single walled culture vessels, you can remove the glass vessel from its
stand as follows:
1. Disassemble or disconnect all equipment, connections and attachment parts
accessible from the outside. Unscrew the screw connections on the lids. Lift it the
lid carefully from the glass vessel.
2. For cleaning in a dishwasher or when replacing, you can lift single walled glass
vessels from out of the stand.
7.1.3 Stirrer Shafts and Motor
Connections | Cleaning and
Maintenance
Work performed by the user is limited to cleaning the stirrer shaft in the case of
contamination from cells and media components and to checking and replacing the
O-rings that seal the stirrer shaft adapter against the cover plate.
Any more extensive disassembly, the replacement of other internal O-rings and
maintenance of the sliding ring seals may only be carried out by qualified service
personnel.
If the stirrer shaft becomes sluggish or blocked, if the motor cannot be activated or if
unusual running noises become audible, report this to the service department.
Spare and Wearing Parts
BB-39121135
O-ring 23.47+2.62, stirrer shaft adapter for UniVessel® 0.5 L,
lid port seal
BB-39121780
O-ring 29.82+2.62, stirrer shaft adapter for UniVessel® 1–10 L,
lid port seal
7.1.3.1 Cleaning the Stirrer
Shafts after Process Completion
1
3
2
82
Appendix
Before cleaning, verify that all O-rings (4) and (8) are intact.
Danger of destroying the stirrer shaft
Stirrer shafts may not be cleaned in laboratory dishwashers as otherwise there is
a danger that the stirrer shaft will be destroyed!
All parts which are not in direct contact with the media during culturing should
only be cleaned and decontaminated superficially. (Wiping with general
laboratory cleaners, e.g. ethanol)!
During the process, the motor coupling (1) must be mounted in such a way that
slight pressure is applied to the V-seal (3) and thereby seals the port (2) located
below it.
Before re-installing the cleaned stirrer shaft, be sure to check the free movement of
the seal ring pair. As depicted in the figure, the spring on the stirrer shaft should be
pressed down for a moment, so that the seal rings are temporarily separated from
one another. This process prevents the seal rings from constantly sticking to one
another, especially in the event that there is considerable foaming during the final
process.
7.1.4 Stirrers | Cleaning and
Maintenance
If overgrown with cells or coated with deposits from culture media residues, the
stirrers can be cleaned with the usual labor cleaning agents as well as mechanically.
In the case of stirrers with adjustable stirring blades, check the fastening of the
stirring blades and the setting of the insertion angle.
Spare and Wearing Parts
Stirrers do not contain wearing parts. Replacement can be ordered using your order
number.
7.1.5. Baffle Insert and Baffles |
Cleaning and Maintenance
Baffle inserts and|or baffles are mounted at the factory. Removing and installing is
only necessary for cleaning or if you want to check and|or replace the O-ring.
All parts can be mechanically cleaned if overgrown with cells or deposits from media
components.
Spare and Wearing Parts
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
7.1.6 Sparger Pipe with Ring
Sparger | Cleaning and
Maintenance
Detach the sparger pipe from the cover plate to check the O-ring (2) and|or to replace
it or for cleaning or if you want to use a different aeration system.
If the ring sparger is overgrown with deposits from media components from previous
processes and the holes are blocked, you can flush the pipe with water in the counter
current and|or clean the ring sparger with a brush or needle. If necessary, you can
unscrew the set screws (4) out of the ring sparger.
When using cleaning agents, flush the sparger pipe and ring sparger thoroughly with
water to prevent any cleaning agent residues from impacting the subsequent process.
Spare and Wearing Parts
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
Arrange the components as described in t “Figure depicting the sparger pipe and
drawing of the culture vessel”.
Appendix
83
7.1.7 Sparger Pipe with Micro
Sparger | Cleaning and
Maintenance
Detach the sparger pipe from the cover plate to check the O-ring (2) and|or to replace
it or for cleaning or if you want to use a different aeration system.
If the frit (5) is overgrown with deposits from media components from previous
processes and the pores are blocked, you can flush the pipe with water in the counter
current. You can also screw off the frit and clean it with a brush or in the ultrasound
bath.
When using cleaning agents, flush the sparger pipe and frit thoroughly with water to
prevent any cleaning agent residues from impacting the subsequent process.
Spare and Wearing Parts
BB-38399120
Spare frit (5)
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
Arrange the components as described in t “Figure depicting the sparger pipe and
drawing of the culture vessel”.
7.1.8. Sparger Insert |
Inspection, Cleaning and
Maintenance of Culture Vessels
If bubbles bead up from the membrane when gassing or during the pressure test,
this indicates that there are cracks or holes. You can try to repair the membrane
with the silicone glue. Otherwise, you must replace the silicone membrane.
The sparger insert is factory-installed in the culture vessel, if included in the
equipment ordered. If delivered separate, the membrane can be fastened to the holder
or packaged separately.
Membrane assembly, removal from and | or installation in the culture vessel are
required,
– when the silicone membrane is mounted for the first time or has to be replaced
later
– for cleaning or retrofitting the culture vessel to a different gassing device
– when the O-ring in the adapter needs checking and|or replacing.
Mount a membrane as shown below. Before installation in the culture vessel, you can
perform a pressure-hold testing in a vessel and|or in the culture vessel with water
(at max. 1.3 barg).
Replacing the silicone membrane:
1. Unscrew the damaged membrane from the holder.
2. Insert the new membrane onto the hose nozzle on the holder from underneath
while at the same time wrapping it around the braces. The notches ensure that the
membrane is wound and equal distances.
3. Insert the membrane end onto the hose nozzle at the top of the holder. Secure the
connections to the nozzles with hose clamps.
The membrane may neither be seated too taut nor too loose on the holder. It is seated
too tight if it is pressed flat against the holder and too loose if it slips from the notches
when pressurized.
84
UniVessel®
Spare and Wearing Parts
BB-39971449
Silicone membrane 3 +0.35, minimum order 10 m, shorten to
required hose length:
y Sparger insert 1 L: 2.7 m
y Sparger insert 2 L: 5.2 m
y Sparger insert 5 L: 10.4 m
y Sparger insert 10 L: 16 m
BB-39971414
Silicone hose 3.2 +1.6, for connecting sparger insert to ports in
the cover plate
y 1 +200 mm (cut to 2+100 mm)
BB-39820181
Silicone glue
BB-8810079
Membrane filter Midisart® 2000
Upon request
Pressure controller t “Drawing of the vessel”, “Parts lists” for
the culture vessels
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
BB-38336975
Cover plate adapter for tubing connection of gas supply and
outlet
Arrange the components as described in t “Figure showing installation of the sparger
insert”.
7.1.9 Universal Adapter |
Cleaning and Maintenance
y Check any adapters used in previous processes, particularly the inner tube, and
clean it if soiled.
y Check the O-ring (3) and replace if damaged.
Spare and Wearing Parts
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
7.1.10 Inoculation Ports (Septa)
| Cleaning and Maintenance
Replace the inoculation membrane prior to each new process.
Check the O-rings and replace if necessary.
Spare and Wearing Parts
BB-39220672
Inoculation membrane made of latex (self-sealing after piercing
and removal of the syringe | inoculation needle)
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
7.1.11 Dip Tube for Adding
Media or Harvesting | Cleaning
and Maintenance
Check the inner tube and clean if media components from previous processes adhere
to it. Check and replace O-rings (3) and (8) if necessary.
Spare and Wearing Parts
BB-39120945
Wearing parts kit “Culture vessels"
Height Adjustable Dip Tube Only
BB-38337630
Ports for the adapter
BB-39507521
Locking ring 6 +0.7 DIN 471-A2 (7)
BB-38337649
Clamping cone (5)
BB-38336820
Screw cap (6)
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
Arrangement: t “Figures showing the harvesting | sampling tube” and | or
“Assignment of lid ports”.
UniVessel®
85
7.1.12 Spinfilters | Cleaning
and Maintenance
Possible biohazard in the case of contaminated spinfilters!
Observe the safety guidelines for handling with contaminated equipment. In such
cases, we recommend to autoclave first before dismounting the culture vessel
and removing the spinfilter.
As long as included with the order, the spinfilter is mounted at the factory:
– Removal and installation is necessary for cleaning the spinfilter
– For retrofitting to other sampling systems the spinfilter can be removed
– Sampling from the permeate chamber of the spinfilter is carried out using the
dip tube SFF:
– Removal and installation is necessary for cleaning the spinfilter.
Cleaning the Spinfilter:
1. You can clean the spinfilter in the laboratory dishwasher. If the mesh is excessively
coated with media residues or cell fragments, you can clean the exterior
mechanically, e.g. with a brush.
2. You can incubate the spinfilter in 0.5 m NaOH overnight at 80°C. You can
neutralize it with 3% H3PO4 afterwards. Next, rinse carefully with deionized
water.
3. You can clean the spinfilter in the ultrasound bath if necessary.
If you use cleaning agents, you must pay attention to their biological compatibility
and carefully rinse all parts with deionized water after cleaning.
Spare and Wearing Parts
Upon request
O-ring (4), spinfilter – stirrer shaft for UniVessel® 1 L
BB-39120953
O-ring (4), spinfilter – stirrer shaft for UniVessel® 2 L
9.25 + 1.78 EPDM/FDA
BB-39121011
O-ring (4), spinfilter – stirrer shaft for UniVessel® 5 L
14.00 +1.78 EPDM/FDA
BB-39120830
O-ring (4), spinfilter – stirrer shaft for UniVessel® 10 L
15.60 +1.78 EPDM/FDA
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
Component Arrangement: t “Figure below showing spinfilter."
7.1.13 Draft Tube | Cleaning
and Maintenance
Removing and installing is only necessary for cleaning or if you want to check and | or
replace the O-ring.
All parts can be mechanically cleaned if overgrown with cells or deposits from media
components.
Spare and Wearing Parts
BB-39120945
O-ring, installation adapter – lid port, 7.65 +1.78 (EPDM/FDA)
86
Appendix
7.1.14 Dummy Plugs |
Cleaning and Maintenance
Check O-rings and gaskets every time you prepare the culture vessel for a process.
Clean dirty gaskets. Replace if damaged or at regular maintenance intervals, as
dictated by the process.
Do not reuse gaskets if they exhibit pressure marks or hairline cracks and|or if
the material is porous.
Spare and Wearing Parts
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
BB-39120830
O-ring for dummy plugs d 19, 15.6 +1.78, EPDM
Component Arrangement: t “Figure below showing dummy plugs."
7.1.15 Pt-100 Temperature
Sensor | Cleaning and
Maintenance
Check, clean and | or replace O-rings every time you prepare the culture vessel for a
process.
The part of the Pt-100 which is immersed in the medium, may be coated by cells,
cell fragments or media residues. You can clean the shaft with a brush.
If the measurement and control system transmits a warning about defects on the
Pt-100, please follow the error analysis and troubleshooting instructions in
t “Documentation on the measurement and control system”.
Frequently, there is a cable break in the connecting cable. Contact the service
department.
Spare and Wearing Parts
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
7.1.16 Antifoam and Level
Probes | Cleaning and
Maintenance
y Check the O-rings. The O-ring around the outer adapter seals the adapter against
the lid port, the O-ring inside the adapter, the probe against the adapter. Clean
the O-rings and|or replace them if they are damaged.
The part of the electrode which is immersed in the medium, may be coated by cells,
cell fragments or media residues. You can clean the shaft with a brush.
If the measurement and control system transmits a warning about defects, please
follow the error analysis and troubleshooting instructions in t “Documentation on
the measurement and control system”.
Spare and Wearing Parts
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
BB-39120821
O-ring (inner adapter) 6.07 +1.78 EPDM (not shown)
7.1.17 Quadruple Addition
Port Adapter | Cleaning and
Maintenance
y Check the addition port adapter used for previous processes, particularly the inner
tube, and clean it in case of contamination.
y Check the O-ring (3) and replace if damaged.
Spare and Wearing Parts
BB-39120830
O-ring, lid port adapter, 15.6+1.78 (EPDM/FDA)
Appendix
87
7.1.18 STT Quick Connector |
Cleaning and Maintenance
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
BB-39120945
You will find information about the spare and wearing parts kit
in Section t “7.4.3"
7.1.19 pH Electrode | Cleaning
and Maintenance
Cleaning
Do not allow soiled pH electrodes to lay around for longer periods.
When media residues dry and harden on the diaphragm, this can make pH
electrodes unusable.
1. Rinse the tip and diaphragm of the electrode with distilled water after each use.
2. Afterwards, you can store the electrode in demineralized water for a short period
of time.
3. In the case of extended storage, make sure that the sensor tip and the diaphragm
of the pH electrode are immersed in 3m KCL solution. For this purpose, you can fill
the protective cap supplied with the solution and put it on the sensor tip.
Follow the manufacturer’s instructions for removing deposits from the diaphragm
using thiourea or pepsin hydrochloric acid solution.
Maintenance
pH electrodes are subject to age and wear from the thermic influences of sterilization
or chemical reactions between the media and diaphragm or electrolyte. Even deposits
on the diaphragm, e.g. proteins, can influence its properties. Product lifetimes depend
on the process conditions, see t"Manufacturer’s instructions”.
The function test is restricted to checking the zero point and slope after calibration.
Follow the instructions given in the documents supplied with the electrode.
Symptoms of wear include, among others, poor triggering, reduced slope or zero point
drift. If the pH measurement differs between standing culture media and that of
agitated culture media, this can be an indication for diaphragm fouling.
Spare and Wearing Parts, Consumables
BB-39120872
O-ring 10.77+2.62 EPDM/FDA
88
Appendix
7.1.20 pO2 Electrodes |
Cleaning and Maintenance
Product lifetimes depend on the process conditions, follow the
t “Manufacturer’s instructions”.
Check the function of the pO2 electrode regularly t “Function test”.
After use, you can rinse the sensor membrane with distilled water and store the
electrode in demineralized water for a short period of time.
pO2 electrodes are subject to wear due to thermic influences (sterilization) and | or
chemical influences (reaction of media particles with the membrane or the
electrolyte). Even growth and deposits on the membrane, e.g. of adhering proteins,
can influence the properties.
Spare and Wearing Parts, Consumables
BB-39123243
O-ring 10.77+2.62 EPDM/FDA
y Arrangement of parts (subject to change): t “Figure showing pO2 electrode”.
Function Test of the pO2 Electrode
The sensor function can be tested by measuring the response time. Prior to checking,
the pO2 electrode must be polarized for at least 2 hours, see calibration instructions.
1. Point the electrode towards the sky. Measure the pO2 value and wait for the
measurement to display stability. Note down this value.
2. Take the electrode to an oxygen-free environment (e.g. a chamber gassed with
nitrogen). Wait until the displayed pO2 measurement is stable.
y After changing from air into an oxygen-free atmosphere , the displayed
measurement must change to less than 1.5% of the value measured in air after
five minutes.
In the case of malfunctions, the electrode must be serviced. For that purpose,
replace the electrolyte and | or the membrane cartridge. Follow the manufacturer’s
instructions on t “pO2 electrode”.
7.1.21 Redox Electrode |
Cleaning and Maintenance
y Redox electrodes are subject to age and wear from the thermic influences of
sterilization or chemical reactions between the media and diaphragm or
electrolyte. Even deposits on the diaphragm, e.g. proteins, can influence its
properties.
y The potential lifetime depends on the process. For information on lifetimes,
signs of wear and how to remove deposits on the diaphragm, please refer to the
t “Manufacturer’s instructions”.
1. Check the O-rings and clean and | or replace them if damaged.
2. Do not allow your soiled Redox electrodes to lay around for longer periods.
Once media residues have dried on, they it may not be possible to remove them
and they can make the electrode unusable.
3. After each use, rinse the tip and the diaphragm of the electrode with distilled
water and store them in distilled water for a short period of time.
Appendix
89
Spare and Wearing Parts, Consumables
7.1.22 FUNDALUX® II |
Cleaning and Maintenance
Order no.
Component
BB-34108165
BB-34108122
BB-33010862
BB-38252290
BB-38343967
BB-39120830
BB-39250750
Redox combination electrode 4805-DPAS-SC-K8S/200, UniVessel® 1 L, 2 L
Redox combination electrode 4805-DPAS-SC-K8S/325, UniVessel® 5 L, 10 L
Connecting cable for pH | rH electrodes
Fitting long shaft, for d 19 mm lid port, UniVessel® 1 L
Fitting short shaft, for d 19 mm lid port, UniVessel® 2 L, 5 L, 10 L
O-ring fitting outer – lid port, 15.6 +1.78, EP
Redox (rH) buffer solution, bottle with 250 ml
Danger of uncontrollable spillage of media in the case of high pressure inside the
culture vessel.
For retrofitting, the culture vessel must be without pressure and at ambient
temperature.
When using probes d 25 mm in side nozzles, the vessel must be emptied to below
the nozzle level.
FUNDALUX® turbidity measurement systems to determine optical density can be
purchased optionally. You will find further details in the t “Fundalux® operating
instructions”.
90
Appendix
7.1.23 Exhaust Cooler |
Cleaning and Maintenance
y Check the O-rings and replace if contaminated or damaged.
If you use culture media which tend to foaming or if you have observed increasing
foam development in-process, you should, from time to time, check the inner tube
for soiling. For that purpose, the headpiece can be removed. Clean the inner tube.
Spare and Wearing Parts
BB-39121763
O-ring (2) head piece to exhaust cooler B5, 18.77 +1.78
EPDM | FDA
BB-39120830
O-Ring (1) lid port fitting, 15.6 +1.78, EPDM | FDA
7.1.24 Sterile Filters for Air |
Gas Supply and Exhaust |
Cleaning and Maintenance
1. The filters cannot be cleaned or serviced. Filters which are blocked by moisture
and|or condensate can be carefully dried with clean, dry compressed air. You must
replace filters whenever penetrated by foam or media.
2. You can autoclave the sterile membrane filters multiple times and use them for
multiple processes. The number of uses depends on the operating conditions.
The potential lifetimes can only be determined empirically.
Spare and Wearing Parts
The filters should be replaced completely t “Product | Ordering information”.
7.1.25 Inoculation Ports (Septa)
d 19 mm | Cleaning and
Maintenance
Replace the inoculation membrane prior to each new process. Check the O-ring on
the dummy plug and replace it, if necessary.
Spare and Wearing Parts
BB-39120830
O-ring for dummy plugs (4), 15.6 +1.78 EPDM | FDA
Appendix
91
7.1.26 1-Channel Inoculation Kit |
Cleaning and Maintenance
If inoculation kits have already been used, be sure to test the inner tube and
inoculation needle (1).
1. Remove any adhering residual media.
2. Check the O-ring and replace it in the case of wear, e.g. when it has pressure
marks or it became porous.
3. From time to time, check the fiberglass filling (2) in the sterile sleeve.
– Unscrew the screw cap (3) and the hose nozzle.
– Replace the fiberglass if wet or soiled.
3
O-ring
2
1
1-Channel Inoculation Kit
Spare and Wearing Parts
BB-39120830
O-ring, dummy plug, 15.6 +1.78 EPDM/FDA
7.1.27 Cooling Fingers |
Cleaning and Maintenance
Check regularly if the coolant hoses and O-rings are in perfect working order.
Replace the hoses and|or O-rings if they are damaged.
The cooling water and|or the cooling agent must be free of residues and deposits
deriving from the supply line. Additionally, avoid residues inside the cooling finger
which therefore remains maintenance free.
If overgrown with cells or coated with deposits from culture media residues, the
insertion sleeves can be cleaned using conventional laboratory cleaning agents as
well as mechanically.
Spare and Wearing Parts
BB-39224511
Hose clamp for silicone hose, 4 units (8)
BB-39120830
O-ring, lid port adapter, 15.6+1.78 EPDM/FDA (12)
7.1.28 Bypass Sampler |
Cleaning and Maintenance
1. The inoculation membrane in the membrane holder is self-sealing and can be
punctured multiple times. It must be replaced prior to each process.
2. Check if the O-ring on the 2-channel screw socket and the silicon hoses are in
perfect working order and replace them if they are damaged.
Spare and Wearing Parts
BB-39120830
O-ring, adapter lid plate, 15.6+1.78 EPDM
7.1.29 Corrective Solution Bottles |
Cleaning and Maintenance
You can clean the glass bottle and|or polypropylene bottle and stainless steel parts in
conventional laboratory dishwashers using conventional laboratory cleaning agents.
Spare and Wearing Parts
BB-38805014
Gasket for screw cap 250 mL – 500 mL bottle
BB-38805332
Sealing for screw cap 10 L, 20 L and 50 L bottle
BB-39224805
Sterile filter for aeration of 250 mL bottle
BB-39237745
Sterile filter for aeration of 500 mL and 1000 mL bottle
Upon request
Sterile filter for aeration of 10 L, 20 L and 50 L bottle
92
Appendix
7.2 Tempering Medium
Danger of malfunction of the temperature control or leaks in the temperature
control circulation.
Contamination or residual corrosion from the laboratory supply line as well as
limescale can cause functional impairments on the valves, the thermostatic pump
and the hose couplings.
Use clean tap water from the supply line without fouling and deposits.
If necessary, install a suitable prefilter.
(Industrial) process water is usually not sufficient.
When operating the bioreactor in a closed temperature control circulation of
the laboratory, the tempering media should not contain any anticorrosion or
antifreeze agents.
If such media additives are contained, you must ensure that they cannot impair
the functions of the instruments inside the temperature control circulation.
The temperature control system of the bioreactors together with “UniVessel®”
culture vessels is intended for water as tempering medium. To avoid limescale
inside the temperature control circulation and the double walled culture vessels,
the water hardness should be not more than 12° dH.
For converting the data you obtain from your water works, use the table below.
Discolorations of the water, particularly a green discoloration in the double wall
can be an indication for algal formation resulting from the use of unclean water.
Water Hardness Conversion Table
1 mmol/l alkaline earth ions
1 mval/l alkaline earth ions
1° German hardness [°d]
1 ppm CaCO3
1° English hardness [°e]
1° French hardness [°f]
Alkaline
earth ions
[mmol/l]
Alkaline
earth ions
[mval/l]
German
hardness
[°d]
1.00
0.50
0.18
0.01
0.14
0.10
2.00
1.00
0.357
0.020
0.285
0.200
5.50
2.80
1.00
0.056
0.798
0.560
CaCO3
[ppm]
English
Hardness
[°e]
French
Hardness
[°f]
100.00
50.00
17.80
1.00
14.30
10.00
7.02
3.51
1.25
0.0702
1.00
0.702
10.00
5
1.78
0.10
1.43
1.00
Appendix
93
7.3 Overview of Assembly
and Equipment
7.3.1 Equipment Options
The equipment, type, arrangement and amount of lid ports as shown on the pictures of
the culture vessels are examples. Vessels tailored to the customer’s specifications can
entail a change in type, arrangement and number or installation of other components.
7.3.2 General Instructions on
Installing Vessel Equipment
y Dummy plugs can be mounted in the lid ports of the culture vessels upon delivery.
When you need the openings for attachment parts, you will have to remove them
first.
y Installing and refitting the following parts require removal of the lid plate from
the glass vessel:
– Stirring elements (e.g. 6-blade disk impeller, 3-blade segment impeller, special
stirrers)
– Baffle insert|baffles and | or draft tube
– Sparger pipe with ring or micro sparger, sparger insert
– Spinfilter with matching sampling tube and draft tube, as appropriate
– Sampling and harvesting tubing
– Adapter for antifoam and|or level probe, inoculation ports and height adjustable
harvesting tube
– Universal adapter (for headspace aeration and additional vessel inlet ports)
– Dummy plugs for unused d 6 mm lid ports
Prior to assembly of the lid plate, make sure which vessel equipment you need.
After assembly of the lid plate, you can only retrofit parts that can be fastened
externally. Follow the instructions for the parts.
y You will find an overview of the different lid configurations in the following.
A detailed description of the vessel equipment and handling is given in the
sections 3–5 of the manual.
y If attachment parts are intended for certain lid ports, a description will be given.
Otherwise, the arrangement in the lid plate is freely selectable and depends on the
equipment which needs to be accessible for operation during process.
y Equipment modified to the customer’s specifications will not be described
separately as long as it can be handled like comparable standard components.
If necessary, descriptions for customer’s specified equipment will be attached to
the customer’s documentation or will be delivered separately.
y If you have questions concerning handling and want to order additional vessel
equipment, spare or wearing parts, please refer to the order number indicated (or
to the labelling on the drawings, if included with customer-specified equipment).
94
Appendix
7.3.3 Installing New Glass
Vessels
Glass vessels, O-rings and gaskets must be replaced if they are damaged.
Spare and Wearing Parts of the Culture Vessels
Culture vessel model
Spare part
Order no.
0.5 L, double walled
Reaction vessel UniVessel®
Wearing parts kit
BB-39204606
BB-34090415
1 L, double walled
Reaction vessel UniVessel®
Wearing parts kit
BB-39204839
BB-34090423
1 L, single wall
Reaction vessel
Wearing parts kit
BB-39204820
BB-34090422
2 L, double walled
Reaction vessel UniVessel®
Wearing parts kit
BB-39204685
BB-34090425
2 L, single wall
Reaction vessel
Wearing parts kit
BB-39204693
BB-34090424
5 L, double walled
Reaction vessel UniVessel®
Wearing parts kit
BB-39204766
BB-34090427
5 L, single wall
Reaction vessel
Wearing parts kit
BB-39204774
BB-34090426
10 L, double walled
Reaction vessel UniVessel®
Wearing parts kit
BB-39204782
BB-34090429
10 L, single wall
Reaction vessel
Wearing parts kit
BB-39204790
BB-34090428
You will find Information on spare and wearing parts for customer-tailored
culture vessels in the equipment documentation.
Appendix
95
7.3.4 Assembly of Glass Vessels
on the Stand
Make sure not to damage the glass flange and the side nozzles for connecting
the temperature control circulation.
7.3.4.1 Jacketed Culture Vessels
1. Turn the glass vessel upside down and place the glass flange on a clean surface,
e.g. a rubber mat. Fit the stand to the glass vessel without nudging the glass
nozzle.
2. Slightly lift up the stand ring to be able to screw the vessel supports into the
drillings of the stand ring. Place each vessel support with its rubber coating onto
the glass edge, align them to the thread drillings of the stand ring and secure
them.
3. Take both the glass vessel and the stand ring, lift them up together and carefully
turn them 180°. Place the glass vessel on the stand onto your work table.
4. Check the position of the vessel supports. The glass vessel must rest evenly.
5. After retrofitting the lid plate, you can align it to the glass flange and secure it.
7.3.4.2 Single Walled Culture
Vessels
1. Set up the stand at your workplace. Check the vessel supports.
They must be secured carefully to the stand ring with the rubber coating pointing
upwards.
2. Lift the glass vessel carefully into the stand without bumping the stand ring.
The glass flange must rest evenly on the supports of the stand ring.
3. If the glass flange rests unevenly on the supports of the stand ring, loosen its
screw connections in order to adjust the position of the glass vessel.
7.3.5 Tubing Connections on
Double-Walled Glass Vessels
Danger of breakage.
If incorrectly arranged (tubing adapter with plug-in sleeve on the lower vessel
nozzle, adapter with hose coupling above) and connection of the supply line to
the upper nozzle, the double wall will build up pressure and may burst in the case
of unallowed overpressure.
1. Check the hoses and gaskets inside the adapters. Replace them if they are
damaged.
During replacement of hose parts, make sure to fasten the hose clips onto the
hose nozzle and the plug-in sleeve and | or hose coupling so tight that they
cannot move.
2. Attach the tubing adapter for the connection to the thermostatic cycle with the
GL-18 screw caps to the glass nozzle on the double wall t “Drawings of the
culture vessels":
y Hose piece with coupling on the lower nozzle inside the double wall. This coupling
is self-closing. In the case the vessel disconnects from the temperature control
circulation after filling, the double wall prevents tempering media from spilling.
y Insert hose piece with plug-in sleeve on the upper nozzle.
This plug-in sleeve is open. After filling the double wall and during autoclaving,
this enables the tempering media to overflow to achieve pressure compensation
between the double wall and its environment.
3. After filling the double wall and during the process, check if water escapes at the
GL-18 connections. Either the screw connection is loose or the gasket in the cap
is defective. First, screw the GL-18 cap on tighter. If this does not help, replace the
gasket.
96
Appendix
7.3.6 Installing Vessel
Equipment
1. When installing vessel equipment, check all O-rings and gaskets. Only flawless
gaskets guarantee sterile sealing. You can slightly lubricate all gaskets with
silicone grease. That way, you prevent the contact surfaces from getting stuck
and damaged.
2. Install all the parts you need for the process into the culture vessel and | or into
the lid plate from below. If possible, equip the lid plate starting from the middle
outward.
Instructions concerning the arrangement of the parts are given below in the
figures to each lid plate. Further instructions concerning the installation
procedure are given in the relevant section.
All parts which can be externally operated in-process have to be easy to access.
3. Fasten all screws and nuts with an appropriate screw wrench.
4. Place the lid plate on the glass flange. The drillings for the lid screws must be
congruent to the drillings in the stand ring. The O-ring that seals the lid plate
against the glass vessel is not allowed to be altered.
5. Plug the knurled screws through the drillings of the lid plate into the thread at the
stand ring. Tighten the screws carefully in a diagonal pattern until lid plate and
O–ring rest evenly on the glass vessel.
Danger of breakage. If you screw the screw caps unevenly and too tightly,
pressure can occur, causing damage to the glass flange and the culture vessel.
Screw caps can loosen due to thermal expansion caused by heating up and
cooling down in the autoclave. This can impair the sterile seal between lid plate
and glass vessel.
Check the tightness of all cap screws after each autoclaving cycle.
7.3.7 Installation and
Connection of Other
Equipment
y The pH electrode must be calibrated prior to installation in the culture vessel.
After sterilization, calibrate the installed pO2 electrode t “Calibration of pH
electrode and | or pO2 electrode” (in the documentation on the measurement and
control system)
1. Mount all sensors and equipment parts which are to be installed after the lid plate
is secured. Connect the peripheral devices which should be autoclaved with the
culture vessel:
– pH and pO2 electrode
– Level and antifoam probe
– Sterile filter supply air
– Exhaust cooler with exhaust filter
– Inoculation ports (septa)
– Cooling finger (for single walled culture vessels)
– Other adapters
– Connections for corrective solution supply
– Sampling devices
y Detailed information on the attachment parts and connections as well as their
handling is given in section 3.
2. To be able to adjust the level electrode for the intended filling volume to the
proper height, measure and mark the fill level indicated on the culture vessel after
installing all parts.
Appendix
97
Example for assignment of attachment parts to lid ports UniVessel® 0.5 L
N1
N2
N3
N4
N5
N6
N7
N8
N9
N10
N11
Exhaust cooler
Addition port adapter 3+DN4
Pt-100 temperature sensor
pH electrode
Addition port adapter 3+DN4
Antifoam | level probe
pO2 electrode
Harvesting pipe
Sampling probe
Supply air
Drive|motor
Example for assignment of attachment parts to lid ports UniVessel® 1 L
N1
N2
N3
N4
N5
N6
N7
N8
N9
N10
N11
Exhaust cooler
Sparger pipe
Pt-100 temperature sensor
pH electrode
4-way port
Antifoam | level probe
pO2 electrode
Harvesting pipe
Baffle insert
Reserve
Level probe
Example for assignment of attachment parts to lid ports UniVessel® 2 L
N1
N2
N3
N4
N5
N6
N7
N8
N9
N10
N11
N12
N13
N14
98
Appendix
Exhaust cooler
Temperature sensor
Antifoam probe
pH electrode
4-way port
Reserve
Reserve
Level probe
Harvesting pipe
pH electrode
Supply air
Baffle insert
Reserve
Sparger insert
Example for assignment of attachment parts to lid ports UniVessel® 5 L
N1
N2
N3
N4
N5
N6
N7
N8
N9
N10
N11
N12
N13
N14
Exhaust cooler
Temperature sensor
Antifoam probe
pH electrode
4-way port
Reserve
Reserve
Level probe
Harvesting pipe
pH electrode
Supply air
Baffle insert
Reserve
Sparger insert
Example for assignment of attachment parts to lid ports UniVessel® 10 L
N1
N2
N3
N4
N5
N6
N7
N8
N9
N10
N11
N12
N13
N14
N15
N16
Exhaust cooler
Sparger insert
pH electrode
Harvesting pipe
Reserve
Supply air
4-way port
pO2 electrode
Sparger insert
Reserve
Baffle
Reserve
Pt-100 temperature sensor
Level probe
Antifoam probe
Reserve
Appendix
99
7.4 Preparing the Culture
Vessels for the Process
y This section aims to give an overview of the steps. Detailed instructions on starting
up the process are given in the documentation on the bioreactor.
7.4.1 Filling in the Jacket of
Double-Walled Culture Vessels
Danger of breakage.
If arranged improperly (tubing adapter with plug-in sleeve on the lower vessel
nozzle, adapter with hose coupling on the upper) and connection to the supply
line on the upper nozzle, the double wall will build up pressure and may burst.
Not applicable for single walled culture vessels, e.g. BIOSTAT® Aplus.
y To ensure optimum heat transfer, the double wall must be completely filled with
tempering medium. Prior to each sterilization and | or process start-up, check the
filling level and add any media lacking.
1. Connect the culture vessel to the temperature control system (on the basic unit):
– The lower connection on the double wall is the inlet to the culture vessel
– The upper port on the double wall is the return
2. Activate the “Fill thermostat” function t “Operating instructions for the micro
DCU system” or push the button on the temperature control module t “BIOSTAT®
Qplus or BIOSTAT® B-DCU”. You can monitor the filling process at the double wall.
After filling, you can disconnect the tubes and continue preparing the culture
vessel.
y The quick coupling on the lower port closes the inlet to the double wall as long as
the connection to the temperature control system is not connected.
The upper outlet (male connector to the quick coupling) is open. You should not
clamp the tube.
7.4.2 Filling the Culture Vessels
1. Fill the culture media in through one of the lid ports. Important note:
y During sterilization, some of the culture media will evaporate. The loss due to
evaporation is about 100 mL for every 30 min. of sterilization time.
Exact values can only be determined empirically. You can fill in additional media
or you can compensate the loss after autoclaving through the sterile addition of
medium.
2. If the culture media is not autoclavable (heat can denature enclosed proteins and
vitamins, sugar can caramelize), fill in some water (about 200 … 300 mL), at least
the amount that gets vaporized during sterilization.
Avoid empty sterilization of the culture vessels because it does not guarantee
safe sterilization.
100
Appendix
7.4.3 Other Pre-process
Measures to Be Taken
It is not necessary to perform the steps in the shown order. The reasonable order
depends on the circumstances for the process.
7.4.3.1 Mounting the Motor
on the Stirrer Shaft
y The motor is connected to the basic unit via a fixed cable attachment and is ready
for operation after mounted onto the culture vessel.
1. Attach the motor to the stirrer shaft on the culture vessel. If necessary, turn it a
little bit until the coupling socket engages with the coupling piece of the stirrer
shaft.
2. Secure it with the locking screw in the ring groove on the stirrer shaft adapter.
7.4.3.2 Connecting the
Aeration|Gassing
1. Connect the air supply (or gas supply) to the supply air filter.
2. Disconnect the silicone hose to the air supply and the clamp on the culture vessel
and | or release the hose clamp.
y Prior to supplying air or oxygen-containing gas, you have to calibrate the zero
point of the pO2 electrode t “Operating manual on ’Micro DCU System’ or ’DCU
Tower.’”)
7.4.3.3 Connecting the Sensors
1. Remove the protective caps or aluminum foil from the plugs on the sensors.
2. Connect the sensors to the bioreactor at the corresponding plugs using the
connection cables (to the supply unit or to the measurement and control system).
3. After activating the basic unit, check if there are any noticeable malfunctions.
Any possible error messages of the measurement and control system will be
displayed on the operator terminal.
4. For the process, choose the necessary measurement and control functions and
configure the corresponding parameters. t “Operating manual on ’Micro DCU
System’ or ’DCU Tower’” Before you activate gassing with air or oxygenated gas,
perform the zero point calibration of the pO2 electrode.
7.4.3.4 Connecting the
Corrective Solution Supply
1. Attach the connection tubing of the corrective solution bottles to the
corresponding pumps.
2. Pump the corrective solution by activating the pumps, until the tubes are filled:
– Using the hand switch on the pump provided you are using pumps with
switches
– In the main menu or the controller operator screen of the control unit, if the
pump has no switch
y If you do not take the empty volumes of the tubes into account or compensate for
them, the proportioning and|or feed volume of the corrective solution will not be
determined correctly.
Appendix
101
7.4.3.5 Connecting the
Tempering System
Double Walled Culture Vessels
Overpressure can destroy the double wall of the culture vessel.
The lower double wall port serves as an inlet for the tempering medium,
the upper port as a return from the double wall to the control unit (to the
temperature control module).
Do not block the return. Operate the temperature control circulation only at
ambient pressure.
1. Check if the cooling water supply on the laboratory side is set to the pressure
stated in the t “P&I Diagram” for the bioreactor and|or set this pressure before
activating the temperature control system:
– Laboratory-side pressure on bioreactors without internal pressure regulation
in the temperature control circulation: max. 1.5 barg.
– Laboratory-side pressure on bioreactors with internal pressure regulation to
1.5 barg in the temperature control circulation: up to max. 2 barg.
2. Connect the supply and return tubing of the thermostatic system.
3. Connect the cooling water intake and return to the exhaust cooler.
Other Notes
y Only when the double wall is filled completely is the temperature transmission
ratio at an optimum. If there is a lack of media, push the “Fill thermostat” button
or activate the corresponding filling function in the operating menu after
switching on the control unit (or the temperature control module).
y The lowest temperature possible in the culture vessel is about 8°C above the
cooling water temperature. For processes with low temperatures, you need a
laboratory cooling device or an external cooling circulation.
Single Walled Culture Vessels with Heating Jacket
Malfunction or damage of the heating jacket if the wrong power supply is used.
Use the connection of the control unit (of the temperature control module) only,
avoid using any other power connection in the laboratory. That is the only way to
ensure the correct power supply of the heating jacket as well as the correct
activation and deactivation of heating and cooling.
y Connect the heating jacket and the cooling water supply and return to the
cooling finger.
7.4.3.6 Process Start
1. Check the preconfigured performance data on the operator terminal of the basic
unit or the measurement and control system and set them for the process,
if necessary, e.g.:
– Operating temperature
– rpms
– Target values, upper and lower pH limit value for pH control
– Target values, limit values and operating mode of the pO2 control
– Reaction thresholds for antifoam and level probe
– Corrective solution and substrate supply, media sampling
(e.g. level control)
2. Inoculate the culture vessel with the prepared inoculation culture.
7.5 Assembly and Operation
of Special Vessels
y In bioreactors with customized culture vessels, the description of assembly,
equipment and operation may be delivered separately or filed in the “Technical
documentation” folder supplied together with the bioreactor.
102
Appendix
7.6 Decontamination Declaration
Defective devices and accessory parts can be sent to Sartorius Stedim Systems GmbH
for warranty claims and service. This shipment must include a decontamination
declaration noting which media, microorganisms and|or cells the device or
components have been in contact with and what type of disinfection and|or
decontamination it has undergone. The recipient must be able to view the
decontamination declaration before unpacking the goods.
A copy of the decontamination declaration for sending devices to Sartorius Stedim
Systems GmbH is included in this appendix or available as a separate document upon
request. Fur further documentation, please contact your representative or Sartorius
Stedim Systems GmbH directly.
Appendix
103
For your notes
104
Notes
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Sartorius Stedim
Systems GmbH
Service
Department
Robert-Bosch-Str."
5–7 78 . 7
,
34302 Guxhagen
89:
*
Germany
Decontamination Declaration
105
Sartorius Stedim Biotech GmbH
August-Spindler-Str. 11
37079 Goettingen, Germany
Phone +49.551.308.0
Fax +49.551.308.32 89
www.sartorius-stedim.com
Copyright by
Sartorius Stedim Biotech GmbH,
Goettingen, Germany.
All rights reserved. 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.
The status of the information,
specifications and illustrations
in this manual is indicated by
the date given below.
Sartorius Stedim Biotech GmbH
reserves the right to make
changes to the technology, features, specifications and design
of the equipment without notice.
Status:
February 2012,
Sartorius Stedim Biotech GmbH,
Goettingen, Germany
Printed in Germany on paper that has
been bleached without any use of chlorine
W_UniVessel
Publication No.: SBT6018-e12021
Ver. 02 | 2012