Operating Manual_DASbox Manual DASbox
User Manual: Operating-Manual_DASbox
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e-Bedienungsanleitung DASbox® Mini Bioreactor System Operating Manual Copyright © 2014 DASGIP Information and Process Technology GmbH, Jülich. All rights reserved, including graphics an images. No part of this publication may be reproduced without the prior permission of the copyright owner. Eppendorf® and the Eppendorf logo are registered trademarks of Eppendorf AG, Hamburg, Germany. DASGIP®, DASbox®, DASware® and BioBLU® are registered trademarks of DASGIP Information and Process Technology GmbH. Teflon® is a registered trademark of DuPont. C-Flex® is a registered trademark of Saint-Gobain Performance Plastics. Bioprene® and Marprene® are registered trademarks of Watson-Marlow Limited. Registered trademarks are not marked in all cases with ® or TM in this manual. 78000148-03.1/112014 Table of contents DASbox® Mini Bioreactor System English (EN) Table of contents 1 User instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.1 How to use this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2.1 Danger symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2.2 Danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.3 Symbols used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.4 Abbreviations used and definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.5 Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 5 6 6 6 7 7 9 2 Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.1 Main illustration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.1.1 DASbox® System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.1.2 DASbox® Bioreactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2 Delivery package of the DASbox® Standard System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3 Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3.1 Technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3.2 DASGIP® Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3.3 Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 11 11 13 14 14 14 15 15 3 General safety instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1 Intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.2 User profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.3 Notes on product liability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.4 Warnings for intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.5 Warning symbols on devices and accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 17 17 18 18 24 4 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.1 Selecting the location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.2 Ports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.2.1 Mains connection unit and communication ports . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.2.2 DASbox® Base Unit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.2.3 MP8-pHpO multi pump and sensor module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.3 Installing the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.3.1 Establishing the electrical supply and communication . . . . . . . . . . . . . . . . . . . . . . . . 4.3.2 Connecting off-gas condensers, overhead drives and temperature sensors . . . . . . . . 4.3.3 Connecting Sensors and cables to the MP8-pHpO module . . . . . . . . . . . . . . . . . . . . . 4.3.4 Connecting the gas supply . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 27 28 28 30 33 34 35 38 39 40 5 Hardware and operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.1 Gassing, temperature control and agitation - the DASbox® Base Unit . . . . . . . . . . . . . . . . . . . 5.1.1 Gas mixing and gassing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.1.2 Temperature control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.1.3 Agitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.2 Off-gas condensers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.2.1 Assembly of the off-gas condensers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.2.2 Operation of the off-gas condensers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.2.3 Deicing the off-gas condensers - Deice function . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3 Addition of media - the MP8-pHpO module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 41 42 43 43 44 45 47 48 49 1 2 Table of contents DASbox® Mini Bioreactor System English (EN) 5.3.1 Feed tubing and feed ranges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3.2 Continuous and pulsed feeding operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3.3 Inserting pump head tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3.4 Calibrating pump head tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3.5 Sterilizing feed tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3.6 Manual operation of the pumps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Monitoring - the MP8-pHpO module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.4.1 Sterilizing sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.4.2 Calibrating sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.4.3 pH sensors - handling instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.4.4 DO sensors (Clark type) - handling instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.4.5 DO sensors for single-use bioreactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.4.6 Redox sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . DASbox® Storage and autoclavable cage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.5.1 DASbox® Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.5.2 LCD monitor display principle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.5.3 Autoclavable Carrier System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . DASbox® Bioreactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.6.1 Typical head plate assignments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.6.2 Glass reactor assembly instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.6.3 Assembling the overhead stirrer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cultivation – a typical workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 51 52 54 54 55 57 57 58 60 61 63 64 65 65 65 67 69 70 71 72 73 6 Software - DASGIP Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1 Short overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1.1 Login . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1.2 Overview of Operations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1.3 New Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1.4 Functional Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2 Functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.1 Gassing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.2 Temperature control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.3 Agitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.4 Addition of media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.5 Monitoring . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75 75 75 76 77 77 90 90 91 91 92 93 7 Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.1 Cleaning and decontamination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.1.1 Surfaces, modules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.1.2 Cleaning feed tubing and pump tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.1.3 Decontamination before service or shipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.2 Service . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.3 Additional information on care and maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.3.1 DO sensors (Clark type) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.3.2 pH sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.3.3 MP8-pHpO module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 95 95 95 96 97 97 97 98 99 8 Transport, storage and disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 8.1 Transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 8.2 Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 5.4 5.5 5.6 5.7 Table of contents DASbox® Mini Bioreactor System English (EN) 8.3 8.4 9 Disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103 General return delivery information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.1 Ambient conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2 DASbox® System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2.1 Power supply . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2.2 Weight / dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2.3 Bioreactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2.4 Agitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2.5 Gassing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2.6 Dispensing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2.7 Monitoring and control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.3 Ports - specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.3.1 Connection assignments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.3.2 Communication ports (DTP in/out, AUX) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.3.3 RS485 (optional) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.3.4 Sensor connectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105 105 105 105 106 106 107 107 107 107 109 110 110 110 111 10 Ordering information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.1 DASbox base unit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.2 System / cables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.3 MP8-pH4pO4 multi pump and sensor module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.4 DASbox Bioreactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.5 Single-Use Vessels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.6 Glass Reactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113 113 114 115 117 117 117 3 4 Table of contents DASbox® Mini Bioreactor System English (EN) User instructions DASbox® Mini Bioreactor System English (EN) 1 User instructions This operating manual describes the design of a DASbox System and explains the handling and operation of its system elements. The DASbox System is also referred to in short as the DASbox in this operating manual. 1.1 How to use this manual Make sure to carefully read this operating manual before using the DASbox for the first time. This operating manual is part of the product. It must always be kept to hand at the site of use. If this operating manual is lost, please request a replacement: bioprocess-support@eppendorf.de or phone: +49(0)2461-980-0. This operating manual applies to two versions of the DASbox: • Version for cell culture with max. 5 sl/h gassing (for CO2 max. 3.5 sl/h*) • Version for microbiology with max. 25 sl/h gassing (for CO2 max. 18 sl/h*). *Note: Due to the physical properties of the gas, the same maximum flow rates cannot be guaranteed for CO2 as for the other gases with which the DASbox System is operated. The versions for cell culture and microbiology also differ in respect to the type of associated bioreactors. Differences between the versions are indicated in the corresponding sections of the operating manual. Other applicable documents The DASbox System is controlled via the DASGIP Control software. Therefore, the complete operating manual for the DASbox System consists of: • this operating manual • the User Manual DASGIP Control Special information on handling DASbox Bioreactors can be found in the following documents: • BioBLU 0.3c/f Single-Use Vessel Instructions for Use • DASGIP Bioreactors. 5 6 User instructions DASbox® Mini Bioreactor System English (EN) 1.2 Danger symbols and danger levels 1.2.1 Danger symbols The warning symbols used in this operating manual have the following meanings: General hazard point Biologically active substances Risk of electric shock Aggressive chemicals Oxidizing substances Explosive substances Hot surfaces Heavy loads Material damage 1.2.2 Danger levels The severity of the danger is indicated by means of signal words. Signal word Severity of danger DANGER Failure to observe this safety information will lead to serious injuries or death. WARNING Failure to observe this safety information may lead to serious injuries or death. CAUTION Failure to observe this safety information may lead to minor or moderate injuries. NOTICE Failure to observe this safety information may lead to product or material damage. User instructions DASbox® Mini Bioreactor System English (EN) 1.3 Symbols used Depiction Meaning You are requested to perform an action. 1. ... 2. ... Perform these actions in the sequence described. • List Useful information 1.4 Abbreviations used and definitions AS Ampere slow CF4/CF6/CF12 4/6/12 mm Compression Fitting COM port Communications port D/OD/ID Diameter/outer diameter/inner diameter DO Dissolved oxygen DoE Design of experiments DTP DASGIP Tiger Protocol; a transmission protocol for the communication with DASGIP modules (devices) via serial interfaces FDA Food and Drug Administration IEC International Electrotechnical Commission LED Light emitting diode 7 8 User instructions DASbox® Mini Bioreactor System English (EN) PA Polyamide PDF Portable Document Format PG (PG13.5) Steel conduit thread PS Polystyrene PTFE Polytetrafluoroethylene; DuPont brand name Teflon® QbD Quality by Design rH Relative humidity RD Redox rpm Rotations/revolutions per minute RTD Resistance temperature detector Rx Receiver SUB-D/D-SUB D-Subminiature; connector type for data Tx Transmitter USB Universal Serial Bus User instructions DASbox® Mini Bioreactor System English (EN) 1.5 Glossary Cleaning-in-place (CIP) A method for the stationary cleaning of process plants or plant parts; in this case, a cleaning procedure for feed tubing without being moved. DASbox® The proper term for the Base Unit of the DASbox System (Chapter 2.1.1). It is often also used as the short name for a complete DASbox System. DASbox® System The term for the sum of all the individual components (DASbox Base Unit and modules) belonging to a DASbox System. DTP network Several DASGIP modules that are connected to one another for communication via serial interfaces. Headspace See: Overlay Level The option of integrating conductivity sensors which can be used for level or foam control. Module A term for a DASGIP device that is used as a component in the DASbox System (examples include the multi pump module or the sensor module). Overlay (Also Headspace); The process of adding media above the surface of the liquid. The media (gas, liquids) are added to the headspace of the bioreactor. Push-in connector/socket A plug-in connection system that enables supply lines to be easily connected and separated. Stirring element A stirrer construction in the bioreactor; a general term used to describe the sum of all components. Submerged The process of adding media below the surface of the liquid. The media (liquids, gas) are added directly into the culture medium. Thermowell Thermometer protection tube. A tube-shaped fixture for the integration of a Pt100 RTD temperature sensor into the bioreactor. Top ring A silicone ring around the edge of a well in the DASbox. Designed to provide protection against broken glass due to impact when handling glass bioreactors. 9 10 User instructions DASbox® Mini Bioreactor System English (EN) Product description DASbox® Mini Bioreactor System English (EN) 2 Product description The DASbox is a unique parallel mini bioreactor system suitable for fermentation in microbiology and cell culture applications. Requiring only minimum lab space, the DASbox provides the functionality of fully equipped industrial bioreactors. 2.1 Main illustration 2.1.1 DASbox® System Fig. 2-1: General view of the DASbox® System 1 DASbox Storage Unit Storage compartments for accessories Monitor for displaying current process values 2 Multi pump and sensor module MP8-pHpO Multi pumps for media feeding Sensor ports for the control of process values: pH, DO, level or pH, DO, redox 3 DASbox Base Unit (Incl. MX4/4-3 gas mixing module) Gas supply, gas mixing Temperature control, agitation 4 Wells for DASbox Bioreactors Wells suitable for BioBLU 0.3c/f Single-Use Vessels and DASbox Glass Reactors with overhead stirrer and off-gas condensation 5 Autoclavable Carrier Removeable carrier for reservoir bottles 6 Gas outlets Push-in ports for gas outlets (overlay/submerged) 11 12 Product description DASbox® Mini Bioreactor System English (EN) Fig. 2-2: Rear view of the DASbox® System 1 DASbox Storage Storage compartments for accessories Port for connecting the monitor 2 Multi pump and sensor module MP8-pHpO Sensor ports for pH, DO, redox, level Pt100 temperature sensor port Communications ports 3 DASbox Base Unit Gas inlets Analog interfaces (optional) Pt100 temperature sensors Overhead drives (for glass reactors or single-use vessels) Off-gas condensers (for glass reactors or single-use vessels) Communications ports Port for connecting the monitor Note: Detailed information on the connection elements at the rear of the system can be found in chapter 4.2, Ports. Product description DASbox® Mini Bioreactor System English (EN) 2.1.2 DASbox® Bioreactors DASbox® Glass Reactors Fig. 2-3: BioBLU® 0.3c/f Single-Use Vessels DASbox® Bioreactors for microbiology and cell culture < 1 Overhead motor For glass reactors or single-use vessels 2 Off-gas condenser with Peltier element For glass reactors or single-use vessels 3a Head plate (GLS80) for glass reactors Ports: 5x Pg13.5 and 4x 4 mm pipe ports Flexible installation of equipment options using DASGIP Compression Fittings 3b Head plate with integrated ports For example, for sensors, addition of liquids, gassing, sampling etc.; magnetic coupling for overhead motor 4a DASGIP Glass Vessel (GLS80) Flat bottom vessel with an outer diameter of 70 mm 4b Reactor vessel with integrated fixtures For example, impellers for cell culture or microbiology, thermowell, port for DO sensor, pipe port 5 Agitation Impellers for microbiology: Rushton type Impellers for cell culture: pitched blade (single-use)/ Marine type (glass reactors) Note: DASbox Glass Reactors and BioBLU 0.3c/f Single-Use Vessels are each available as cell culture and microbiology versions. 13 14 Product description DASbox® Mini Bioreactor System English (EN) 2.2 Delivery package of the DASbox® Standard System The delivery package of a DASbox System depends on the respective configuration and can therefore vary greatly. Please refer to the corresponding delivery note for full details of the scope of your system. The delivery package of a DASbox System always includes the following system components: • DASbox Base Unit with 5 sl/ or 25 sl/h gassing • MP8-pHpO multi pump and sensor module as a combination module with DASbox Storage and Monitor • Process computer with monitor, keyboard and mouse • 4 DASbox Bioreactors (glass reactors or single-use vessels) • Sensors, condensers, overhead drives, tubing for liquids and gases, media and reservoir vessels All of the required cables, connectors and multiple socket strips are included in the delivery package. • Refer to the chapter 10, Ordering information. This chapter contains a list of accessory parts for DASbox Systems. • The multi pump and sensor module is optionally available with level or redox measurement (correct product designation: MP8-PH4PO4LS or MP8-PH4PO4RD4S). 2.3 Features 2.3.1 Technology DASbox System The DASbox features advanced temperature and agitation control and allows for accurate monitoring and control of pH, DO and level. Mass flow controllers control the individual gas supply to the bioreactors. The modular design of the DASbox System allows several systems to be stacked together. The DASbox System can be extended to consist of up to 24 parallel operating bioreactors with a minimum use of space. A parallel system of 24 units requires only 1.8 m of lab bench space. DASbox Bioreactors The DASbox can be operated with glass reactors or single-use bioreactors (BioBLU 0.3c/f Single-Use Vessel). Both reactor types are each available as cell culture or microbiology versions and support applications with a volume range of approximately 60 - 250 ml. • Glass reactors: 60 - 250 ml • Single-use vessels: 65 - 250 ml (microbiology); 100 - 250 ml (cell culture). Product description DASbox® Mini Bioreactor System English (EN) Overhead-operated mini impellers are used for the agitation of the DASbox Bioreactors. • Glass reactors: Rushton type (microbiology) or marine type (cell culture) • Single-use vessels: Rushton type (microbiology) or pitched blade (cell culture) Gases can either be supplied submerged or via the headspace of the bioreactor (overlay). In addition, an integrated splitter function in the DASbox Base Unit enables individual gas types to be routed directly into the medium (submerged) or to the headspace. Off-gas condensers with Peltier elements are available for exhaust gas routing. 2.3.2 DASGIP® Control The DASbox is exclusively controlled via the DASGIP Control software. The software allows for flexible configuration of fermentation processes with online processing of set-point profiles and enhanced automation options via trigger functions. Parallel calibration procedures and preconfigured cleaning-in-place (CIP) procedures help with the preparation and follow-up of your processes. During the fermentation process, numerous intervention options are available via the intuitive process view. DASware® options The DASGIP DASware Bioprocess Software Suite offers a range of flexible and intelligent solutions for the implementation of key Quality by Design (QbD) aspects of the current FDA's Process Validation Guidance, thus accelerating the development of bioprocess operations. In particular, DASware design, a tool for the application of statistical Design of Experiment (DoE), and DASware discover, a comprehensive and user-friendly software for data and information management, are designed to make working with the DASbox even easier. 2.3.3 Applications With the above-mentioned features and mini bioreactors with a minimum working volume, the DASbox is the optimal tool for advanced process development. Other typical applications include screening of cells and cloning, media optimization and statistical Design of Experiment (DoE) applications. 15 16 Product description DASbox® Mini Bioreactor System English (EN) General safety instructions DASbox® Mini Bioreactor System English (EN) 3 General safety instructions This section contains important information for the intended use of the DASbox System. 3.1 Intended use The DASbox System is designed for the cultivation of cells and microorganisms in biotechnological laboratories. Typical areas in which the DASbox System is used include research, process development and quality control in different biotechnology fields (e.g., industrial, pharmaceutical, biotechnology). Only original accessory parts may be used for operating the DASbox System. WARNING! Not approved for use in medical diagnostics or therapy! The DASbox System has not been developed for applications in medical diagnosis or therapy and must not be used in such applications. It is not a medical device within the meaning of Directive 93/42/EEC. WARNING! Risk of electrical hazard when wet. The DASbox System is neither splash-proof nor suitable for outdoor use. The DASbox System is intended for indoor use only. 3.2 User profile The DASbox System must only be operated by qualified personnel. The qualified personnel must have received training for the system described here which has been organized by an Eppendorf Bioprocess Center service technician. The qualified personnel must have carefully read this operating manual. 17 18 General safety instructions DASbox® Mini Bioreactor System English (EN) 3.3 Notes on product liability The operating safety of the DASbox System may be compromised in the cases outlined below. In such cases, the owner is responsible for assuming liability associated with material damage or personal injury claims of any sort. • Commissioning and using the DASbox System without observing the notes and instructions provided in this operating manual and the other applicable documents. • Improper use. • Using accessory parts that are not recommended by DASGIP. • Operating the DASbox System outside of the application limits described in this operating manual. • Maintenance or repair of the system by persons who have not been authorized to do so by Eppendorf AG or DASGIP. • In the case of unauthorized modifications to the DASbox System. 3.4 Warnings for intended use WARNING! Lethal voltages inside the device. Make sure that the housings of the devices belonging to the DASbox System are always closed and undamaged to ensure that no parts inside the devices can be touched inadvertently. Never open the housings of the devices. Avoid the ingress of liquid into the inside of the devices. The devices may only be opened by service personal who have been authorized to carry out such work by DASGIP or Eppendorf. WARNING! Risk of electric shock from damage to devices or mains power cables. Do not switch on any devices belonging to the DASbox System if visibly damaged. Only use devices that have been properly installed or repaired. Disconnect the device from the mains power supply in the event of danger. Only replace damaged parts with original spare parts. Access to the mains power switch must not be blocked. WARNING! Risk from incorrect supply voltage Only connect the devices belonging to the DASbox System to power sources that meet the electrical specifications on the back of the devices. Only use sockets with a protective earth (PE) conductor and a suitable mains power cable. General safety instructions DASbox® Mini Bioreactor System English (EN) WARNING! Electrical hazard if protective grounding is missing. Each device within the DASbox System is connected via a cable with a protective grounding conductor to the mains power supply. Make sure that your mains power supply system has a functional protective ground contact for each device and that it is properly connected to the protective ground contact on the power cable of the device. WARNING! Electrical hazard. Mains isolating device must be freely accessible. In an emergency, it must be possible to quickly unplug the power supply cables of all devices within the DASbox System. When installing the system, ensure that each of the power supply cables of the DASbox System are freely accessible at one of the two ends. WARNING! Electrical hazard when replacing safety fuses in the device. The device safety fuses are located in the mains connection unit at the back of each device. Disconnect the plug on the device from the power supply before replacing device safety fuses. Only use fuses of the following type: Dimensions: 5x20 mm; rated voltage at least 250 V, characteristic "slow-blow". nominal current 2.5 A (MP8-pHpO); 10 A (DASbox Base Unit). Observe the information on the rear panel of the device. DANGER! Risk of explosion. Do not operate the DASbox System in rooms where work is being carried out with explosive substances. Do not use the DASbox System to process any explosive or highly reactive substances. Do not use the DASbox System to process any substances which could create an explosive atmosphere. 19 20 General safety instructions DASbox® Mini Bioreactor System English (EN) CAUTION! Risk of personal injury and material damage when lifting and carrying heavy loads. The DASbox is heavy. Lifting and carrying the device can lead to back injuries. The hooks at the front and the grip at the back of the DASbox are not handles. They are only designed for moving the DASbox on telescopic rails or for lifting the lid during service work. NEVER use the hooks and the grip on the DASbox as handles. The DASbox must be transported by least two people. Lift the DASbox by reaching under the device. Use a transport aid (such as a dolly) to transport the device over longer distances. WARNING! Danger to health from pathogenic germs or infectious liquids. When handling pathogenic germs and infectious liquids, observe the national regulations, the biological safety level of your laboratory, the safety data sheets, and the manufacturer's application notes. Wear personal protective equipment (PPE). WARNING! Danger to health from aggressive chemicals. Observe the safety data sheets of all substances used (e.g., pH adjustment agent). Wear personal protective equipment (PPE). WARNING! Danger from oxidizing or explosive gases and gas mixtures Gas mixtures in specific ratios can be explosive. Determine and observe the upper and lower explosion limits (UEL; LEL) of the gases used. Also consider the possible production of gas during the cultivation process. Do not use the DASbox to process gases within the explosion limits. WARNING! Danger due to oxygen enriched atmospheres. Even a slightly oxygen-enriched atmosphere increases the risk of materials igniting more easily and burning more intensely. Oxygen-enriched hair or clothing can catch fire and result in serious injury or death. There is a risk of self-ignition in case of contact with oils and greases. Only use oil-free compressed air for operating the DASbox. Keep all parts which come into contact with oxygen free from oils and greases. Always make sure that no excess oxygen can increase the ambient concentration to a dangerous level. Regularly check the fitting and tightness of the gassing tubes to the bioreactors. General safety instructions DASbox® Mini Bioreactor System English (EN) WARNING! Maximum operating pressure of 0.5 bar. Exceeding the limit may result in burst risk. DASbox Bioreactors are designed for a maximum operating pressure of 0.5 bar (relative overpressure). Do not operate the bioreactors with a higher pressure. Never block the outlet filters. Observe the maximum inlet pressure of 0.5 bar for all connected gases. Do not connect the gases without a pressure reducer. Wear personal protective equipment (PPE) while handling gassed bioreactors. CAUTION! Risk of explosion due to damaged glass vessels. Hairline cracks, fissures or deep scratches in the glass can endanger the stability of glass reactors. This may pose a risk of explosion! Splinters can fly out in an uncontrolled manner. Inspect for visible signs of damage and do not use damaged glass reactors. Wear protective goggles in the vicinity of gassed bioreactors. CAUTION! Danger from hot surfaces. Please note that the wells for holding the reactors of the DASbox can become hot. G WARNING! Danger from uncontrolled flying parts. In the event of an increase in pressure in the bioreactor, inadequately secured sensors can be ejected from the reactor and become dangerous projectiles. Check the secure fitting of all sensors prior to the start of a fermentation. CAUTION! Danger from rotating parts. When operating overhead operated stirring elements outside of the bioreactor there is a risk of injury and the danger of clothing or hair getting entangled. Do not touch the rotating parts. Only operate overhead operated stirring elements within the bioreactor. 21 22 General safety instructions DASbox® Mini Bioreactor System English (EN) CAUTION! Danger from squirting of dispensing media. If blockages occur on the outlet side of the feed line, this may result in a significant increase in pressure within the supply line due to the pumping capacity of the peristaltic pump. There is the potential risk of aggressive liquids escaping or being squirted out. Regularly check the feed tubing for blockages (kinks, encrustations, etc.). Wear protective goggles and protective clothing. CAUTION! Danger from liquids and gases leaking. Inadequately fastened or incorrectly fitted Luer connectors can result in leaks. Leaked dispensing media can cause encrustation between the DASbox Base Unit and the bioreactor. Check that all Luer connectors are straight and securely attached before each use. NOTICE! Observe the ambient conditions. Observe the permissible ambient conditions when installing and operating the DASbox. Before using the DASbox at high temperatures and high air humidity, contact DASGIP to check system suitability. CAUTION! Do not cover the ventilation slots. Always ensure sufficient convection to allow heat to dissipate. Make sure that the ventilation grating of the system components is not covered. CAUTION! Do not confuse connecting sockets! Observe the connection values. When connecting accessory parts, observe the labeling on the connectors and the specified connection values. When connecting sensors, observe the input limits specified in the manual. CAUTION! Risk of product damage when connecting gases to the DASbox. Particles passing through the gas inlets into the interior of the DASbox can damage valves and flow sensors. Forcibly detaching the connected gas tubing may cause plastic shavings that can get inside the DASbox. Do not connect the gases without a particle filter. Use the supplied disassembly tool to disconnect the push-in connectors. General safety instructions DASbox® Mini Bioreactor System English (EN) CAUTION! Risk of product damage. Do not disconnect any connectors during operation. Devices and accessory parts may be damaged. Do not disconnect any connectors during system operation. CAUTION! Risk of product damage due to increased top-heaviness of bioreactors with an off-gas condenser and a stirring drive. The installation of an off-gas condenser and a stirring drive greatly increases the top-heaviness of DASbox Bioreactors. The reactors may tip over easily. Before assembling accessory parts, ensure the reactor is stable by positioning it in a well in the DASbox. CAUTION! Risk of product damage during the assembly and storage of Pt 100 temperature sensors. The Pt 100 temperature sensors can be bent by improper assembly and storage. In particular, the matt part at the lower end of the sensor can easily break. Carefully guide the sensor into the thermowell. Store unused temperature sensors in a protective covering or similar or in the designated holders provided in the DASbox Storage Unit. CAUTION! Risk of product damage when handling DO sensors for DASbox Single-Use Reactors. The DO sensors for DASbox Single-Use Reactors can be damaged by improper handling. Avoid exerting mechanical stress on the sensor tip. Store unused sensors in the storage box. CAUTION! Risk of collision with rotating stirring elements Rotating stirrer impellers can damage sensors and other fixtures in the bioreactor (this may include broken glass and scratch marks). Make sure that all stirring elements and fixtures are mounted correctly. CAUTION! Risk of damage to device from improper use of cleaning agents. Do not use any aggressive, corrosive or abrasive disinfectants or cleaning agents. If you have any questions about the compatibility of cleaning agents and disinfectants or their ingredients, please contact a service technician at the Eppendorf Bioprocess Center. 23 24 General safety instructions DASbox® Mini Bioreactor System English (EN) 3.5 Warning symbols on devices and accessories Abb. 3-1: General view of the epMotion Fig. 3-1: Warning symbols on the DASbox® System 1 WARNING! Observe the connection values. Observe the information on the connection values provided in the operating manual. 2 CAUTION! Consult user manual. For operation conditions consult user manual 3 CAUTION! Hot surfaces. Please note that the wells for bioreactors can become hot. General safety instructions DASbox® Mini Bioreactor System English (EN) Fig. 3-2: 1 Warning symbols on the DASbox® accessories CAUTION! Hot surfaces. The surfaces of the off-gas condensers and stirring drives can become hot after a longer operating time. 25 26 General safety instructions DASbox® Mini Bioreactor System English (EN) Installation DASbox® Mini Bioreactor System English (EN) 4 Installation The DASbox System is delivered in a transport-safe wooden box. The system must be unpacked and installed by Eppendorf Bioprocess Center service technicians. Have the wooden box transported as close as possible to the future installation site. Do not unpack the box after delivery. Check the packaging for external damage only. In the case of visible damage to the outer packaging, inform your contact person at the Eppendorf Bioprocess Center. Only unpack the box together with a service technician. This procedure enables the technician to check the equipment for any hidden transport damage. If possible keep the padded packaging for future transport or shipment. 4.1 Selecting the location Information on ambient conditions, system dimensions and weights can be found in the technical data (chapter 9). WARNING! Risk of danger from electrical current. The DASbox is suitable for indoor installation only! Select the location for the device according to the following criteria: Location • The ambient conditions match the specifications in the technical data. • The location is dry, clean and free of dust. • The convection of air at the rear of the device must be sufficient to ensure heat can dissipate. • The location is protected from direct sunlight. • The location is not next to heat sources, such as heaters or drier compartments. • The device can be safely and easily operated at this location. • When the device is in operation, the device mains power switch and the mains isolating device (e.g., FI safety switch) must be freely accessible. Workplace • The lab bench is stable and designed to accommodate the weight of the DASbox System. 27 28 Installation DASbox® Mini Bioreactor System English (EN) • The lab bench has a horizontal, plane work surface. • In addition to the system, the lab bench also offers enough space for performing work necessary during the completion of cultivation processes. Electrical connections • 2 mains power connections for a DASbox System with 4 to 8 bioreactors. • Mains power connections: 100-240 VAC (no manual switching required), 50/60 Hz. Gas supply The compressed air and all gases that are connected to the DASbox System must meet the following criteria: • Compressed air: oil-free, dry and free of particles. • Gases: clean, free of particles, dry, non-condensing. CAUTION! Risk of damage to device from incorrect gas quality. Only gases that meet all the above-mentioned criteria may be used for operating the DASbox System. 4.2 Ports 4.2.1 Mains connection unit and communication ports The mains connection unit and communication ports (serial interfaces) are identical for all DASbox devices. Therefore, the details of these connection units are described here first. Installation DASbox® Mini Bioreactor System English (EN) Mains connection unit Fig. 4-1: Communication ports Mains connection unit and communication ports 1 Mains disconnect switch of the device The on and off positions are marked according to standard: I = ON O = OFF WARNING The mains disconnect switch of the device must always be freely accessible. 2 Mains connection IEC power connector for connecting a power cord 100-240 VAC, 50/60 Hz. 3 Fuse holder Fuse holder for holding two 20 mm x 5 mm microfuses. DASbox Base Unit: 2 x 10 AT MP8-pHpO: 2 x 2.5 AT. 4 AUX SUB D9M / RS232 plug for connecting additional devices and potential free alarm contacts. 5 "DTP out" SUB D9M / RS232 plug for integrating DTP-capable add-on modules (e.g., GA4, MP8) to a DTP network. 6 "DTP in" SUB D9F / RS232 connector for communication with the process computer. This port is either connected directly with the process computer, or alternatively with the DTP Out output of another DTP-capable device. 7 "Rx/Tx" (DTP communication display) Gray = no signal Green = receive Orange = transmit Red = communication fault 8 "State" (DTP state indicator) Gray = DTP in/out ports enabled Green = RS485 ports enabled NOTICE! Do not confuse the "RS485" ports with the Ethernet ports as the design is the same. 9)*RJ45 connectors for a RS485 network Potential-free (*assignment optional) 29 30 Installation DASbox® Mini Bioreactor System English (EN) 4.2.2 Fig. 4-2: DASbox® Base Unit Ports at the rear of the DASbox® Base Unit 1 Sticker with important information DASbox version, gas flow, part number, serial number, CE marking, contact information 7 LCD Monitor Port for the LCD monitor integrated in DASbox Storage. 2 Pt 100 1-4 for temperature sensors Pt 100 temperature sensors for measurement in the bioreactors. 8 DTP port for internal system communication The DTP ports (COM Ports) of the DASbox modules are connected with serial DTP cables. 3 Condenser 1-4 for off-gas condensers Off-gas condensers (typically 10 W per condenser) 9 External power supply Mains connection unit, Fig. 4-1 4 Drives 1-4 for overhead motors Overhead motors für DASbox Bioreactors (Glass or Single-Use Bioreactors) 5 Grip Used to open the DASbox for service purposes. Not suitable for carrying purposes. 6 Gas ports and *optional external interfaces For an explanation see Fig. 6-2 10 USB port for external communication This port is connected with a USB cable to a USB port on the process computer. 11 Internal power supply Stas 200 type ports for the internal power supply of the system. 12 Ventilation grating For removing heat from the inside of the device. Do not cover the ventilation grating. * Further information about specifications of ports can be found in chapter 9, Technical data. Installation DASbox® Mini Bioreactor System English (EN) CAUTION! Risk of damage to device. When connecting DASbox components, observe the labeling on the connecting sockets and the specified connection values. Don’t connect the plugs of the overhead motors with the connecting-sockets of the off-gas condensers. The design of the plugs is similar, but not identic. Observe the input limits specified in the manual. Do not disconnect any plugs during system operation. Do not cover the ventilation grating. Gas supply Fig. 4-3: DASbox Base Unit - ports for the gas supply 1* External I/O (optional) External interfaces; option for analog system contol. 2 Gas IN (gas inlets) N2,Air, O2,CO2 Gas inlets for nitrogen, compressed air, oxygen and carbon dioxide. 6 mm push-in connector for connecting gas tubing. To protect the sensors inside the device, particle filters are integrated by default into the gas tubing on the inlet side. 3 Overlay (overlay gas outlets) Outlets of channels 1-4 for gas types which are to be directed into the headspace of the bioreactor (4 mm push-in connector). 4 Submerged (submerged gas outlets) Outlets of channels 1-4 for gas types which are to be directed into the medium (4 mm push-in connector). To protect the sensors inside the device, nonreturn valves have been integrated here to prevent medium from flowing back through the gassing tube. * Further information about specifications of ports can be found in chapter 9, Technical data. < 31 32 Installation DASbox® Mini Bioreactor System English (EN) WARNING! Risk of explosion if oxygen comes into contact with greases and oils. There is a risk of self-ignition if oxygen comes into contact with oils and greases. Only use oil-free compressed air for operating the DASbox. Keep all parts which come into contact with oxygen free from oils and greases. CAUTION! Risk of damage to device if the DASbox is operated without a particle filter. Even if the connected gases have a very high purity grade, a discharge of particles from gas bottles or pipelines cannot be excluded. Particles can be the cause of operational changes in electronic mass flow sensors or leaks in valves. To avoid such errors, particle filters are integrated into the gas tubing on the inlet side. Do not operate the DASbox System without a particle filter. CAUTION! Risk of damage to device from moist compressed air or gases. Moisture can damage the sensors inside the DASbox. Only use dry, non-condensing gases and compressed air for operating the DASbox System. WARNING! Danger from overpressure. Maximum operating pressure of 0.5 bar. DASbox Bioreactors are designed for a maximum operating pressure of 0.5 bar (relative overpressure). Observe the maximum inlet pressure of 0.5 bar for all connected gases. Do not connect the gases without a pressure reducer. Installation DASbox® Mini Bioreactor System English (EN) 4.2.3 Fig. 4-4: MP8-pHpO multi pump and sensor module Rear of the multi pump and sensor unit of the DASbox® 1 DTP ports for internal system communication Serial ports (COM ports) for internal communication among the DASGIP modules. The Serial ports of the DASbox modules are connected by serial cables (DTP cables). 2 pH 1-4 for pH sensors Measuring range: pH 0 – pH 14 (± 0.01 pH) (at a sensor temperature of max. 67°C) 3 pO2 1-4 for DO sensors Measuring range DO: 0 – 500 % DO Measuring range Temperature (NTC): -10°C – 140°C (± 0.2 K) 4 level 1-4 for conductivity sensors (optional: for redox sensors) Conductivity measurement for foam or level control Input limits: 0 – 20 mS Measurement of the redox potential Input limits: -2000 – 2000 mV (± 1 mV) 5 "Pt100" temperature sensor Pt 100 temperature sensor for automatic temperature compensation during calibration and measurement. Input limits: 90 – 160 Ohm Measuring range: -25°C – 116°C (± 1 °C) 6 LCD Monitor Port for the LCD monitor integrated in DASbox Storage. 7 Serial number When contacting DASGIP, please always provide the serial number of the device. 8 Mains connection unit Power supply, mains power switch and fuse holder (see chapter 4.2.1). The mains power connection is connected to the internal mains power supply of the base unit. 9 Ventilation slots Ventilation slots for removing heat from the inside of the device. Always ensure the flow of exhaust air is unrestricted. * Further information about specifications of ports can be found in chapter 9, Technical data. 33 34 Installation DASbox® Mini Bioreactor System English (EN) CAUTION! Risk of damage to device from incorrect connections. When connecting DASbox components, observe the labeling on the connecting sockets and the specified connection values. Observe the input limits specified in the manual. Do not disconnect any plugs during system operation. 4.3 Installing the device CAUTION! Risk of damage to device. The DASbox may only be installed and commissioned by skilled personnel authorized by DASGIP. Please contact the Service Department before making any changes to your system. Refer to the following instructions on connecting components belonging to the DASbox System. Also refer to the information on the connection elements provided in the chapters 4.2.1 to 4.2.3. Installation DASbox® Mini Bioreactor System English (EN) 4.3.1 Establishing the electrical supply and communication First establish all the connections needed to ensure the power supply and to secure communication between the devices. Electrical supply First make sure that all mains power switches are in the "O" (=OFF) position. For the external power supply, use the supplied power cord to connect the power connector of the base unit to a mains socket (1). Then establish the internal power supply of the system: Connect one of the Stas 200 plugs on the DASbox Base Unit to the mains power connection of the MP8-pHpO module (2). *Optional add-on modules: Either connect an add-on module (e.g., MP8 or GA4) via the second Stas 200 plug to the DASbox Base Unit or connect it with an additional power connector to a mains power socket. Use the adapter cable with a Stas 200 (Hirschmann) plug. Lock the Stas 200 plugs by folding down the metal brackets. WARNING! Risk of electric shock. Never use any devices if the power cord or plug is damaged. 35 36 Installation DASbox® Mini Bioreactor System English (EN) Communication (establishing the DTP network) Next establish the connections for device communication. Within a DASbox System, the individual modules are connected in series with each other starting from the base unit using DTP cables (1). The end is formed by a terminator connector (2). Connect the DTP port on the base unit (no. 8 in Fig. 4-2) to the "DTP in" port on the MP8-pHpO module (1). *Optional: If an additional DASGIP module should be integrated, connect the "DTP out" connector on the MP8-pHpO module to the "DTP in" input on the additional module. Provide the "DTP out" output on the MP8-pHpO module (*optional on the additional module) with the terminator‐ connector (2). Note: To ensure stable data transfer, each "DTP out" output of the last device is provided with a terminator connector. For external communication, connect the USB port "PC" on the base unit (no. 10 in Fig. 4-2) to a free USB port on the process computer (3). Use the supplied USB cable for this purpose. Installation DASbox® Mini Bioreactor System English (EN) Connecting the LCD monitor Connect the monitor port at the rear of DASbox Storage to the "LCD" port on the DASbox Base Unit (1). Use the supplied monitor cable for this purpose. 37 38 Installation DASbox® Mini Bioreactor System English (EN) 4.3.2 Connecting off-gas condensers, overhead drives and temperature sensors Now connect all the elements to the DASbox Base Unit which are housed in the storage compartments of DASbox Storage: • Overhead motors (1), • Actuators for off-gas condensers (2), • Temperature sensors (3). Position the actuators for off-gas condensers, overhead motors and the temperature sensors as shown in the adjacent figure in the storage compartments of the DASbox Base Unit. Guide all the cables from front to back through the storage compartment. Now connect the temperature sensors (1) , off-gas condensers (2), and overhead drives (3) for the reactors 1 - 4 to the base unit. Note: Observe the number marks on the cables. NOTICE! Possible risk of damage to connectors! The connectors for the actuators have 8 pins (1); the connectors for the overhead drives have 7 pins (2). They are otherwise identical in construction so that the overhead drives are also suitable for the connectors of the off-gas condensers. Plug and socket connectors must not be confused. Installation DASbox® Mini Bioreactor System English (EN) 4.3.3 Connecting Sensors and cables to the MP8-pHpO module Next, connect the following sensor cables to the MP8-pHpO module: • Pt100 temperature sensor for temperature compensation during pH calibration (1). • 4x pH cable (2) • 4x DO cable (3) • or optionally 4x DO sensors for single-use vessels (3a) (DO sensor for DASGIP D 4.7 mm, L 162 mm) • 4x cable for conductivity sensors for the level option (4) • or optionally 4x cable for redox sensors (4a) Guide the sensor cables from front to back through the cable compartment of the DASbox Storage Unit. Connect the cables according to the labeling to the connecting sockets at the rear of the MP8-pHpO module: Pt100 Sensor (1), pH cables 1-4 (2), DO cables 1-4 or DO sensors 1-4 for single-use vessels (3) cables 1-4 for the level option or redox cables 1-4 (4). Note: Observe the number marks on the cables. 39 40 Installation DASbox® Mini Bioreactor System English (EN) 4.3.4 Connecting the gas supply Connect the gas tubing of the four inlet gases to the "Gas IN" gas inlets at the rear of the DASbox Base Unit. NOTICE! Risk of damage to device. Do not connect inlet gases without a particle filter and a pressure reducer. There is a maximum inlet pressure of 0.5 bar. Connect the gassing tubes to the "Overlay" and "Submerged" outputs next to the wells for bioreactors. Note: Insert the supply lines into the push-in ports applying slight pressure, until you can feel them engage. Loosen the gassing tubes To loosen the gassing tubes, use the provided disassembly tool for push-in ports. Pull the gas tube out while pressing down the outer ring of the push-in port. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5 Hardware and operation 5.1 Gassing, temperature control and agitation - the DASbox® Base Unit Fig. 5-1: Functions of the DASbox® Base Unit 1 Agitation Individual agitation with overhead motors for glass reactors and single-use vessels. 4 Autoclavable Carrier Removable storage facility for up to 8 reservoir bottles with media (see chapter 5.5). 2 Off-gas condensers Suitable for for glass reactors and single-use vessels, see chapter 5.2. 5 Temperature control Wells for 4 DASbox Bioreactors, suitable for glass reactors and single-use vessels. Individual temperature control, liquid free using Peltier elements. 3 Gassing Gas mixing system with integrated splitter function: "Overlay" and "Submerged" gas outputs next to each bioreactor well. 6 Top rings Silicone impact protectors designed to prevent broken glass when handling DASbox Glass Reactors. 41 42 Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.1.1 Gas mixing and gassing Gas mixing system The gas mixing system integrated in the DASbox is fully mass flow controlled and supports software configurable gassing schemes. It has 4 gas inlets as well as 4 device channels, each of which can be assigned to a bioreactor. The gas inlets are supplied by default with N2, Air, O2 and CO2. Each device channel is equipped with two outlets for submerged or overlay gassing (see below "Integrated splitter function"). 4 x 4 independently operating mass flow controllers for each gas and each channel allow for continuous mixing of individual gas compositions. Variants For different applications the DASbox Gas Mixing System is available in two versions: • Microbiology: 0 .. 30 sl/h (for CO2 max. 18 sl/h*) • Cell culture: 0 .. 5 sl/h (for CO2 max. 3.5 sl/h*) • *Due to the physical properties of the gas, the same maximum flow rates cannot be guaranteed for CO2 as for the other gases with which the DASbox Gas Mixing System is operated. • The accuracy of flow values can not be guaranteed above the maximum flow rates. • Nonreturn valves are integrated inside the device on the gas output side. They prevent medium from flowing back into the interior of the device. Integrated splitter function The DASbox Gas Mixing System features a splitter function. For each channel it can be determined which of the connected input gases should be directed through proportional valves to one of the two output channels ("Overlay" or Submerged"). Defining the supply of gas types either to the headspace of the bioreactor or directly into the medium is thus possible. The assignment of the gas types to the gas outlets is defined via the DASGIP Control software. Find more information in the User Manual DASGIP Control. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.1.2 Temperature control The DASbox is equipped with a quick and accurate temperature control for each individual bioreactor. It operates in a range of 10 – 60°C (at an ambient temperature of 25°C). Pt 100 temperature sensors (class A RTD) ensure high temperature measurement accuracy. The cooling system is completely free of liquid. An additional supply of coolant is not required. Find more information in chapter 6.2.2 and in the User Manual DASGIP Control. 5.1.3 Agitation The highly accurate DASbox agitation control enables a working range of between 20 and 2500 rpm. For single-use vessels, the upper speed limit is 2000 rpm. In DASbox Systems for cell culture applications, the maximal permissible speed is limited to 500 rpm within the DASGIP Control software. Depending on the type of application, DASbox Bioreactors are equipped with Rushton type impellers or Marine type respectively pitched blade impellers. The glass reactors have miniaturized industry standard lip seal assemblies. The impellers are driven by brushless overhead drives offering long service life and minimal operating noise. Support for online calculation of the stirrer torque provides feedback on culture viscosity. Notice! Risk of damage to overhead drive. The overhead drives are not waterproof. Do not immerse the overhead drives in liquid. 43 44 Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.2 Off-gas condensers The DASbox features special miniaturized off-gas condensers operated with Peltier elements. They allow liquid-free cooling and heating without additional supply of coolant. The DASbox Off-Gas Condenser is characterized by easy handling. Magnetic connection and disconnection is used to enable or disable cooling operation. A thermally insulated construction minimizes the formation of condensation on the outside of the exhaust gas line. Component parts Abb. 5-2: Component parts of the DASbox® Off-Gas Condensers for glass reactors and single-use vessels 1 Condenser adapter for glass reactors Adapter plate with exhaust gas line. Version with 12 mm gas pipe for compression fittings (12 mm, CF12). 2 Condenser adapter for single-use vessels Adapterplate with exhaust gas line . Version with an exhaust tube guide for singleuse vessels. 3 Exhaust gas line insulation For glass reactors and single-use vessels (not autoclavable). 4 Temperature control actuator For use with glass reactors and single-use vessels. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.2.1 Assembly of the off-gas condensers Off-gas condensers for glass reactors Install the condenser adapter using a 12 mm Compression Fitting (CF12) in the bioreactor head plate. The adapter plate has to be turned outwards. Press the exhaust gas line insulation onto the condenser adapter. Slide it as far as it will go onto the exhaust gas line tube on the adapter plate. NOTICE! Insulation is not autoclavable. The exhaust gas line insulation is not temperature-resistant and must be removed for autoclaving. Slide the temperature control actuator onto to the adapter plate from above. If necessary, bend the silicone tubing with the outlet filter slightly to the side in the process. If it is positioned correctly, actuator operation is automatically enabled via an integrated magnetic switch. Slide the actuator down from the adapter plate to disable operation. Also refer to the chapter 5.2.2. 45 46 Hardware and operation DASbox® Mini Bioreactor System English (EN) Off-gas condensers for single-use-vessels Carefully insert the condenser adapter into the holder provided in the head plate of the BioBLU 0.3c/f Single-Use Vessel. NOTICE! Risk of material damage. If force is applied during assembly, the lever action of the adapter may cause cracks to form in the holder of the head plate. Press the exhaust gas tubing of the BioBLU 0.3c/f Single-Use Vessel into the groove on the condenser adapter. Press the tube insulation onto the condenser adapter as shown and then push it down to the stop. Slide the actuator onto the plate of the adapter from above. If it is positioned correctly, actuator operation is automatically enabled via an integrated magnetic switch. Slide the actuator down from the adapter plate to disable operation. Also refer to the chapter 5.2.2. CAUTION! Increased top-heaviness. The bioreactors may tip over easily (see next page). Hardware and operation DASbox® Mini Bioreactor System English (EN) CAUTION! Increased top-heaviness of bioreactors with off-gas condensers. The installation of off-gas condensers greatly increases the top-heaviness of single-use bioreactors in particular. The bioreactors may tip over easily. Before assembling the off-gas condensers, position the reactors in the wells of the DASbox Base Unit. 5.2.2 Operation of the off-gas condensers Note: Activate the off-gas condensers BEFORE gassing is started via the software. The condenser must have reached its operating temperature before exhaust filter blockages can be prevented. Slide the temperature control actuator on to the adapter plate from above. If it is positioned correctly, the actuator is automatically enabled via an integrated magnetic switch. Remove the actuator from the adapter plate to disable condenser operation. NOTICE! Ventilator in operation. The ventilator of the fan rotates at a very high speed! Avoid contact with accessory parts (e.g., Pt100 temperature sensors). As soon as the DASGIP Control software is started, the operating status of the connected off-gas condensers is displayed on the LCD monitor. A blue cooling symbol next to the unit concerned indicates the current operation of the respective off-gas condenser. 47 48 Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.2.3 Deicing the off-gas condensers - Deice function DASbox Off-Gas Condensers always operate at maximum cooling capacity. Longer periods of operation may result in condensate freezing on cooled components. If this happens, the temperature control actuator can no longer be detached from the condenser adapter plate. A setting in the DASGIP Control software therefore ensures that the operation of the Peltier elements is automatically briefly reversed after each process is completed. For five minutes the off-gas condenser is not cooled but heated in order to facilitate the disassembly of the temperature control actuator. The DASGIP Control software also provides the "Deice" function by means of which the off-gas condenser can also be heated during a process run for five minutes. As soon as the actuator is separated from the adapter plate, the control switches back to standby mode. Cooling operation starts again as soon as the actuator and the adapter plate are brought together again or after the short heating period has elapsed. Starting the "Deice" function During the process run, click on the symbol for temperature control T Controller in the process view of the software. The window containing the temperature control parameters is opened. Activate the Deice function by clicking on the Deice button. The actuator briefly switches from cooling to heating mode. The Deice function is automatically stopped afterwards. You can now remove the temperature control actuator from the adapter plate. The condenser heating mode is indicated by a yellow heating symbol next to the unit concerned on the DASbox LCD monitor. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.3 Addition of media - the MP8-pHpO module The DASbox MP8-pHpO Module combines the functions of the DASGIP MP8 Multi Pump Module and the DASGIP pHpO Sensor Module in a single device. This chapter describes the main functions of the MP8 multi pump module. Eight electronic speed-controlled feed pumps ensure the supply of DASbox Bioreactors with liquid media. In standard process configurations, two feeding lines are thus available per bioreactor which enable the continuous addition of nutrients and optimal pH control. By using different pump tube types, a wide range of different feeding rates from 0 to 420 ml/h are possible. The integrated calibration of the individual pumps ensures reproducible results. 5.3.1 Feed tubing and feed ranges DASbox Feed Tubing DASbox Feed Tubing consists of three individual parts: The connection line between the reservoir bottle and the pump is connected via a flexible piece of tubing (pump head tubing) to the connecting line between the pump and the bioreactor. Fig. 5-3: Design of DASbox® Feed Tubing 1 Feed tubing A connecting line between the reservoir bottle and the pump or the pump and the bioreactor. Different lengths are available. Material: PTFE (not resistant to autoclaving) C-Flex (autoclavable) 2 Pump head tubing A flexible piece of tubing that is inserted into the tubing bed of the pump head for pumping liquids. Different inner diameters are available for variable feeding ranges. Material of the pump head tubing: Bioprene or Marprene (both autoclavable) Note: Pump head tubing is a wear part. Replace pump head tubing at regular intervals. 3 Luer connectors Easy connection of reservoir bottles and bioreactors. Quick replacement of worn pump head tubing. 49 50 Hardware and operation DASbox® Mini Bioreactor System English (EN) Pump head tubing For the operation of the DASbox in standard applications, pump head tubing with an inner diameter of 0.5 mm is used. With this type of tube, a feeding range of 1.3 - 39 ml/h is possible. Depending on requirements, other ranges can be covered by varying the pump head tubing used. For information on standard tube types and the corresponding feeding ranges, refer to Fig. 5-4 and the associated table 5-1. Fig. 5-4: Typical feeding rates (F in mL/h) for different pump head tubing diameters (ID in mm) The table shows the minimum and maximum feeding rates that can be achieved belonging to pump head tubing as well as the typical calibration factor FCal. The calibration factor FCal is defined as the number of pump head revolutions per milliliter of the fed medium. The exact value of the factor is determined during the calibration of the pump head tubing. Tab. 5-1: Feeding ranges of different pump head tubes ID x wall [mm] Fmin [ml/h] Fmax [ml/h] FCal [1/ml] 0.25 x 1.0 0.3 9.5 450 0.5 x 1.0 1.3 39 110 1 x 1.0 4.0 122 35 2 x 0.8 14.0 420 10 *The values of the pump head tubes used as a standard are indicated in bold. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.3.2 Continuous and pulsed feeding operation Continuous feeding operation The area limited by the minimum and maximum feeding rates in Fig. 5-4 shows the operating range which allows a continuous feeding operation of the multi pumps. • Continuous feeding operation is indicated by the pump LED remaining constantly lit: Green: clockwise rotation Orange: counterclockwise rotation Pulsed feeding operation As soon as the values for the minimum possible feeding rates drop below the set-points, the pump switches to a pulsed operating mode. In this mode, longer periods of pump inactivity alternate with feeding operation phases. • In this operating mode, the pump LED only remains lit as long as the pump is rotating. • When the pump is stationary, a short flashing in longer intervals indicates that the pump is active and is in the standing phase of the pulsed feeding operating mode. For further clarification of this principle of operation, refer to the typical working ranges of the pumps in the table below. Tab. 5-2: Typical working ranges of the pumps Working range [%] [rpm] Maximum possible speed of the pump head (100%) 71 Upper speed limit of the typical operating range (60%) 43 (upper speed limit) Minimum speed for continuous operation* (3.3%) 2.4 (lower speed limit) *Pulsed feeding operation is activated if the speed drops below the minimum values. 51 52 Hardware and operation DASbox® Mini Bioreactor System English (EN) CAUTION! Risk of damage to pumps by exceeding the upper speed limit! The upper speed limit of the typical pump operating range (60%) must not be permanently exceeded. Permanently exceeding the limit will reduce the service life of the tubing and drive. Use pump head tubing with a larger inner diameter. If the pump is operated permanently in the minimum speed region within an application, we recommend the use of pump head tubing with a smaller inner diameter. This allows feeding accuracy to be improved. 5.3.3 Fig. 5-5: Inserting pump head tubing Design of the pump head 1 Pump guard 4 Tube guide on the left side 2 Tubing bed 5 Pump rotor 3 Tube guide on the right side The pump must be switched off before the pump head tubing is inserted. Follow the instructions below: Hardware and operation DASbox® Mini Bioreactor System English (EN) 1. Open the pump guard. The tubing bed is lifted automatically and the pump head tubing can be loaded. 2. Insert the pump head tubing into the pump tube guide at the bottom left. 3. Route the pump head tubing around the pump rotor and insert it into the tube guide on the right side. In the case of thicker tube types it may be necessary to press the tubing into the tubing bed by applying light pressure with your finger. 4. Grasp the elastic tube at both ends and pull it down slightly. 5. Check the position of the tubing in the tubing bed. The tubing must not be jammed in the tubing bed. 6. Close the pump guard. The tubing bed is lowered and the pump head tubing is pressed gently between tubing bed and pump rotor. CAUTION! Risk of deformation of the pump head tubing. Do not allow pump tubing to be jammed in the tubing bed for prolonged periods. This can lead to permanent deformation and thus to inhomogeneous feeding. If the pump is not used for an extended period, open the pump guard in order to loosen the tubing bed. 53 54 Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.3.4 Calibrating pump head tubing To be able to operate the pumps with defined feeding rates, the pump head tubing must first be calibrated. The DASGIP Control software is used for calibration: During this process, each of the theoretical feeding amounts per time unit are compared with the actual feeding amounts within a preconfigured feeding program, taking into account the density of each media. In addition, a calibration factor FCal [1/ml] is determined, i.e., the number of pump head revolutions per milliliter of the fed medium. For more information, refer to the User Manual DASGIP Control. It describes in detail how to carry out the calibration procedure. 5.3.5 Sterilizing feed tubing DASbox Feed Tubing in microbiology applications is usually sterilized using a cleaning-in-place (CIP) procedure. PTFE supply lines are used in most cases. In cell culture applications, the supply lines are usually autoclaved for sterilization. For this reason, C-Flex® supply lines are used. CIP (cleaning-in-place) procedure The sequential application of ethanol and sodium hydroxide is described in literature as an effective sterilization method, which also exhibits a deactivating effect on spores. (Source: P. Girot, Y. Moroux, X.P. Duteil, C. Nguyen, E. Boschetti; Composite affinity sorbents and their cleaning in place; Journal of chromatography, (1990) 510 213-223.) The DASGIP Control software contains a preconfigured CIP procedure: During this process, the feed tubing is successively rinsed with ethanol (70%) and sodium hydroxide (2 N). This CIP procedure can be customized as needed. Refer to the User Manual DASGIP Control. This manual describes in detail how to carry out the CIP procedure. NOTICE! Limited chemical resistance of C-Flex® tubing! C-Flex® tubing is not suitable for use with alcohols and organic solvents. Do not carry out CIP procedures on this type of feed tubing. Autoclave the supply lines instead. Hardware and operation DASbox® Mini Bioreactor System English (EN) Autoclaving You can also remove C-Flex® feed tubing from the mini pumps for autoclaving. This practice is especially standard in cell culture applications: Autoclave the feed tubing individually or connected with the reservoir bottles for media. Seal the ends of the supply lines, e.g., with the corresponding Luer caps or aluminum foil. CAUTION! Risk of product damage to feed tubing from autoclaving! Only autoclave thoroughly rinsed feed tubing. Media residues can crystallize in the supply lines and cause them to become irreversibly blocked. Do not sterilize PTFE feed tubing by autoclaving. PTFE feed tubing is not resistant to autoclaving. Do not seal feed tubing made of C-Flex® with a hose clip for autoclaving. The C-Flex® material may become irreparably deformed. 5.3.6 Manual operation of the pumps The mini pumps are usually controlled by the DASGIP Control software. The software also offers several options for intervention during the process run. Manually operating the MP8 is possible, but not absolutely necessary. The following sections contain the main possibilities of manual operation: Activating the manual operating mode To switch a pump to manual operating mode, press both arrow keys 1 x short simultaneously. The indicator light (LED) of the pump flashes orange and green alternately. 55 56 Hardware and operation DASbox® Mini Bioreactor System English (EN) Short feeding (< 4 seconds) To perform very short feeding (< 4 seconds), either press the right or left arrow key for the desired seconds, depending on the direction of rotation. • The pump will stop as soon as the button is released. • The LED will flash as long as the pump is running: Clockwise rotation green Counterclockwise rotation orange Continuous feeding Press the right or left arrow key for longer than a few seconds (default 4 s) to activate the continuous feeding operating mode. • In this operating mode, the pump will continue to run after the control button has been released. • Briefly pressing one of the two arrow keys again stops the pump. • The start of continuous feeding is indicated by a short acoustic signal. • The LED will flash as long as the pump is running: Clockwise rotation green Counterclockwise rotation orange Ending the manual operating mode • Briefly pressing both arrow keys simultaneously switches the pump from manual mode back to computer-controlled mode. Note: During the switch to remote control operation, the pump immediately accepts the last commands that were transmitted. The pumps may start immediately! Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.4 Monitoring - the MP8-pHpO module The MP8 - pHpO module combines the functions of a DASGIP MP8 Multi Pump Module and a pHpO sensor module in a single device. This chapter contains important information on the functions of the sensor module and the associated sensors. The pHpO sensor module supports the use of standard pH and DO sensors for the measurement and control of process values. A separate input for a Pt100 temperature sensor allows for automatic temperature compensation during calibration and measurement. In addition, the control of conductivity sensors for level control or as an anti foam probe is supported. As an alternative to the level option, a redox measurement is offered. CAUTION! Risk of sensor damage from improper handling. The sensors supplied with the DASbox are sensitive wet-chemical systems that require proper handling, good care and regular maintenance. Make sure to observe the manufacturer's instructions for the respective sensor types. Please follow the instructions and information in this chapter and in chapter 7.3. 5.4.1 Sterilizing sensors Please observe the following information: • The DASbox sensors which are in direct contact with the culture medium can generally be sterilized by autoclaving. Before using alternative sterilization methods, please observe the instructions of the respective sensor manufacturer. • All Pt100 temperature sensors and the DO sensors for use with single-use bioreactors are not autoclavable. • The sensors used in DASbox Glass Reactors are generally integrated in the bioreactor and autoclaved along with it. • The calibration of pH sensors takes place BEFORE autoclaving (see typical workflow Fig. 5-7). • The calibration of DO sensors only takes place AFTER autoclaving (see typical workflow Fig. 5-7). Add a few drops of water into the bioreactor for autoclaving as the sensors should ideally be autoclaved in a steam atmosphere. 57 58 Hardware and operation DASbox® Mini Bioreactor System English (EN) Seal the connectors of the sensors with the corresponding cap in order to prevent the ingress of moisture. CAUTION! Risk of damage to sensors. Do not autoclave the pH sensors in deionized water under any circumstances. The electrode gel will otherwise bleed out. Information on the use of single-use bioreactors • Autoclavable sensors which are operated in single-use bioreactors can be sterilized in advance and then fitted into the single-use reactor under sterile conditions. For more information, also refer to the BioBLU® 0.3c/f Single-Use Vessel Instructions for Use. 5.4.2 Calibrating sensors The DASGIP Control software contains procedures for carrying out parallel one or two-point calibrations of pH and DO sensors: In the navigation tree on the left side, right-click on the desired calibration entry to open the context menu. Click on Run to start the calibration procedure. Refer to the User Manual DASGIP Control. This manual contains detailed descriptions how to carry out sen‐sor calibrations using the control software. Observe the following information on calibrating sensors: pH sensors - information on calibrating We recommend carrying out a two-point calibration of the pH sensors before autoclaving (see workflow, Fig. 5-7). When carrying out calibration, take into account the desired working pH-range for the intended use. Typically, the buffers pH 4 and pH 7 can be used for calibration. Note: Do not mix up sensor channels. During the cultivation process, the sensors must be used for the MP8-pHpO module channels for which they were previously calibrated. Subsequently mixing up the sensor channels will make it necessary for the calibration to be repeated. After disconnecting the sensors from the sensor module (e.g., for autoclaving purposes), make sure they are reconnected with the correct assignment. Hardware and operation DASbox® Mini Bioreactor System English (EN) DO sensors - information on calibrating The following information also applies to DO sensors for use in single-use bioreactors. • In contrast to the pH sensors, the DO sensors are only calibrated AFTER autoclaving. The slope of the sensor can be influenced by the autoclaving process. • We generally recommend a one-point calibration (at 100 % saturation) immediately before the start of the main procedure (see workflow, Fig. 5-7). • A two-point calibration is only necessary for processes in very low DO ranges or which require a very high level of accuracy. • Calibration must always be carried out BEFORE the culture is inoculated. • Calibration can be carried out in the culture medium. The medium must be in a state of equilibrium with the ambient air, i.e., saturated at 100% with the oxygen of the ambient air. • To achieve the saturation state, set a medium gassing rate, start a light agitation and control the temperature of the medium to the desired operating temperature. • Ensure that operating conditions are constant. • As soon as a constant DO value has been reached, the calibration procedure can be started in the DASGIP Control software. For more information, refer to the User Manual DASGIP Control. Redox sensors - verification of the correct functioning Their correct functioning can be verified with the aid of buffers with defined redox potentials or a Chinon / Hydrochinon redox test. Please follow the instructions in the documentation provided by the sensor manufacturer. The DASGIP Control software features a check procedure which supports such a function test. If you have any questions, contact a technician at the Eppendorf Bioprocess Center. Level Sensors Level sensors do not need to be calibrated. 59 60 Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.4.3 pH sensors - handling instructions Please observe the following instructions for the correct handling of pH sensors to ensure the proper function of the sensors. In addition, always also observe the instructions of the respective sensor manufacturer. General • pH sensors generally have a limited service life. Their aging is essentially accelerated by the number of autoclaving cycles. • The thickness of the source layer of the sensitive glass increases over the course of time, contributing to the inertia of the sensor. The sensor becomes slower. • After several autoclaving cycles, the electrolyte can take on a brown discoloration. This has no effect on the functioning of the sensor. • Signs of contamination of the diaphragm include: • • • • Long response times Unstable measurement signals Strong offset shifts Low sensor slope You can find information on the regeneration of pH sensors in chapter 7.3.2. Correct storage Store the pH sensors upright in KCl solution (Mettler Toledo 3.0 mol/l, Broadley James 3.8 mol/l). You can store the pH sensor in buffer pH 4 for a short period if no KCl solution is available. Rinse the sensor with tap water and pat it gently dry when you remove it from the KCl solution before use. CAUTION! Risk of sensor damage. Do not leave the sensor in the open air for a long period of time. Do not store the sensors in deionized water. The electrode gel will otherwise bleed out. Visual sensor check Before each use of the pH sensors, inspect the sensors for the following: • The plug connector must be dry and free from signs of corrosion and contamination. Otherwise, the transmission of measured values may be interrupted. Hardware and operation DASbox® Mini Bioreactor System English (EN) • The sensor cables must not have any sharp bends or be brittle. • The sensor tip must be completely filled with electrolyte solution. If necessary, shake the sensor gently like a clinical thermometer, so that it fills up. • The pH-sensitive glass of the sensor tip must be free of cracks, scratches or deposits. • Usually, the diaphragm of the pH sensor is white. Gray-black discoloration indicates contamination with sulfide. Refer to the chapter 7.3.2 for information on cleaning and regeneration of pH sensors. Checking the raw signal of the pH Sensors During the calibration process of the pH sensors, pay attention to the raw signal of the sensor as an important indicator of its proper functioning. • Limit values: pH 7 buffer raw 0 mV ± 5 mV pH 4 buffer raw 180 mV ± 10 mV • Regenerate the sensor or replace it if the sensor slope is below 90% after a two-point calibration. Note: The raw value (raw) in mV can be displayed during the calibration procedure by clicking on the pH sensor symbol. For information on carrying out calibration, refer to the User Manual DASGIP Control. 5.4.4 DO sensors (Clark type) - handling instructions Please observe the following instructions for the correct handling of DO sensors to ensure the proper functioning of the sensors. In addition, always also observe the instructions of the respective sensor manufacturer. Visual sensor check Before each calibration and each time the DO sensors are used, check the following points: • The plug connectors must be dry and free from signs of corrosion and contamination. Otherwise, the transmission of measured values may be interrupted. • The sensor cables must not have any sharp bends or be brittle. • Inspect the sensor membrane for visual signs of damage or contamination. • Clean contaminated membranes by wiping with a soft, damp cloth. 61 62 Hardware and operation DASbox® Mini Bioreactor System English (EN) • Replace membranes with strong signs of wear or scratches. • Replacing the membrane body can also be useful in the case of non-calibrated sensors, long response times, unstable measured values or significant drift of measured values. • Replace the electrolyte solution at regular intervals (see chapter 7.3.1). Note: A baggy membrane has no effect on the functioning of the sensor, as long as the membrane is intact. Checking the raw signal of the DO Sensors During the calibration process of the DO sensors, pay attention to the raw signal of the sensor as an important indicator of its proper functioning. • Limit values: 100% DO raw between 50 nA and 100 nA (slope) 0% DO raw 0 up to 3 nA (offset) Polarize The sensors must be allowed to polarize for approximately 10 h before use. For this, connect the sensors to the corresponding connectors on the pHpO sensor module and switch on the device. Note: Polarized sensors may be temporarily disconnected from the sensor module. However, an equally long subsequent polarization time should always be observed afterwards. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.4.5 DO sensors for single-use bioreactors Observe the BioBLU® 0.3c/f Single-Use Vessel Instructions for Use for more detailed instructions on operating the sensors in single-use bioreactors (DO sensor DASGIP D 4.7 mm, L 162 mm). Storage Store the DO sensors for use in single-use bioreactors in the black screw box in a humid atmosphere. Regularly moisten the sponges located in the screw cap with deionized water. Store the sensors in an upright position. CAUTION! Risk of sensor damage. Avoid mechanical stress and handle the sensors with care. Protect the sensors from hard impacts. Do not hold the membrane by the sensor tip unnecessarily and keep it away from sharp objects. The sensors are not autoclavable. Polarization The sensors must be allowed to polarize for approximately 10 h before use. For this, connect the sensors to the corresponding connectors on the pHpO sensor module and switch on the device. Note: Polarized sensors may be temporarily disconnected from the sensor module. However, an equally long subsequent polarization time should always be observed afterwards. Cleaning Rinse off any encrustation or visible contamination on the sensor tip with warm water. Carefully wipe away any stubborn deposits using a soft, lint-free cloth. Do not use any pointed or sharp-edged objects to scrape away contamination. 63 64 Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.4.6 Redox sensors Please observe the following instructions for the correct handling of Redox sensors to ensure the proper functioning of the sensors. In addition, always also observe the instructions of the respective sensor manufacturer. Check the diaphragm of the sensor. It should be bright, white and intact. Check the internal pressure condition and sealing. The sensor must show sufficient inner pressure and an intact sealing according to the manufacturer’s manual. Check the electrolyte colour. A slight brown discoloration is acceptable and caused by normal tear and wear. Run a full two point calibration according to the calibration procedure in the DASGIP control Software Store the sensors in 3M KCl solution , when not in use. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.5 DASbox® Storage and autoclavable cage 5.5.1 DASbox® Storage The storage unit DASbox Storage forms the upper part of the DASbox System. It provides special storage compartments for DASbox accessories: Use of storage compartments • DASbox Storage provides storage space for typical DASbox accessories. 1. Overhead motors 2. Pt100 temperature sensors 3. Off-gas condensers Store all accessory parts that are not needed as shown in the figure. Note: The Pt100 temperature sensors in particular are thus protected against damage due to improper storage or excessive mechanical stress. 5.5.2 LCD monitor display principle The front of DASbox Storage incorporates a LCD monitor for displaying current process information: Stand by • As soon as the DASbox System is switched on, the display "DASbox Not connected" which is shown on the left appears on the monitor. • This display remains active until the DASGIP Control software is started. 65 66 Hardware and operation DASbox® Mini Bioreactor System English (EN) Current process values • After the DASGIP Control software starts, a four-part display appears. • In this display, current process values can be displayed for each of the four reactors of the relevant DASbox System. • As long as no cultivation has been started via the software, missing measured values are represented as lines in front of the unit concerned. • After a control procedure has been started, current values for pH, DO, temperature and speed are displayed for each bioreactor. Identification of bioreactors • If you click on the view of a single reactor during a cultivation process, this is visually highlighted on the LCD monitor of the relevant DASbox. • An enlarged view of the number of the respective bioreactor appears. A blue spot is blinking. • This simplifies the process of assigning the values observed in the software to a specific bioreactor, especially in the case of large plants with several parallel DASbox Systems. • The normal display appears again after approximately 10 seconds. Condenser status • As soon as off-gas condensers are connected to the DASbox, their operating status is shown by means of a small condenser symbol to the right next to the reactor name: blue = cooling operation yellow = heating operation (Deice function) gray = condenser has been disconnected from the actuator red = internal error message Hardware and operation DASbox® Mini Bioreactor System English (EN) Errors and alarms • The LCD monitor allows for the display of errors and alarms in the four-part view: green = process value OK orange = warning (minor deviation from set-point) red = error (significant deviation from set-point) To visually display errors and alarms, activate the alarm function in the control software. Then start the cultivation run by pressing the Inoculation button. Note: For more information on alarm functions, refer to the User Manual DASGIP Control. 5.5.3 Autoclavable Carrier System The accessories for the DASbox include an autoclavable carrier system which consists of 3 main components: 1. Cage for reservoir bottles 2. Adjustable carrying handle 3. Cage for 4 DASbox Bioreactors • All components are made of autoclavable stainless steel. 67 68 Hardware and operation DASbox® Mini Bioreactor System English (EN) At the front of the DASbox Base Unit there are two screws. Hook in the cage for reservoir bottles there to attach it to the DASbox. Remove the cage from the base unit by lifting it slightly and then unhooking it. Connect the cage for reservoir bottles to the cage for bioreactors by hooking it in at the side screws. Loosen the Allen screws on the cage for bioreactors and position the handle in the middle of the cage construction. Attach the carrying handle by tightening the Allen screws. • This ensures that bioreactors and the associated media bottles can be transported safely, e.g., for autoclaving and that they can also be autoclaved in the cage. Telescopic rails for the DASbox Base Unit • Parallel DASbox Systems consisting of three DASboxes or more are generally mounted on rails. • They allow individual base units to be pulled out to the front. • This option allows for easy handling of the bioreactors, e.g., during sampling. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.6 DASbox® Bioreactors Note: The following chapters are excerpts from the User Manual DASGIP Bioreactors. Due to the wide variety of special equipment options for DASbox Glass Reactors, please refer to this User Manual for further information. It contains detailed information and instructions on the integration and installation of the equipment options mentioned here. Please observe the User Manual DASGIP Bioreactors. For single-use vessels, please observe the BioBLU 0.3c/f Single-Use Vessel Instructions for Use. 69 70 Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.6.1 Typical head plate assignments The head plate of the DASbox Bioreactor supports a wide variety of equipment options via 6 x Pg13.5 and 4 x 4 mm pipe ports. The figure shows a typical head plate assignment for microbiology and cell culture applications. : Fig. 5-6: Typical head plate assignment for a DASbox® Bioreactor 1 Lip seal assembly for overhead stirrer 7 dip tube (4 mm) as a sampling option 2 For example, septumholder with rubber sealing 8 Exhaust gas line with or without off-gas condenser Installation of off-gas condenser: 12 mm Compression Fitting Installation of exhaust gas pipe: 6 mm Compression Fitting 3 Short 4 mm pipe ports for addition of media 4 Gas supply via L-sparger (4 mm Compression Fitting) or sparger (4 mm Compression Fitting) 5 DO sensor 6 ports (2mm) for level sensor 9 Thermowell for temperature sensor 10 pH sensor 11 4 mm dip tube for media feeding Optional equipment The following optional equipment is possible: • OD sensor with Pg13.5 • Sensor for level and anti foam control via a 4 mm Compression Fitting Hardware and operation DASbox® Mini Bioreactor System English (EN) Note: You have the option of changing the arrangement of the equipment options in the head plate. However, please note the following hints: Install pH and DO sensors on opposite sides. In this arrangement the sensors can act as additional baffles. Seal all ports that are not used with blind plugs or rubber seals. Refer to the User Manual DASGIP Bioreactors for more information on installing equipment options and using DASGIP Compression Fittings. 5.6.2 Glass reactor assembly instructions Note: Please keep in mind that some equipment options must be installed in the head plate from below. Install these options before closing the reactor. • On the underside of the head plate there is a groove containing an O-ring for sealing. Before each use, check the O-ring for cracks and damage and replace it if there is visible damage. Place the head plate with the seal on the thread of the glass vessel and center it there. Place the screw cap on the head plate and screw it tight onto the glass thread. Note: Place the screw cap on the head plate already before installing the equipment options. It may no longer be possible to slide the screw cap over all the fixtures later. 71 72 Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.6.3 Assembling the overhead stirrer • The lip seal assembly (2) of the overhead stirrer of DASbox Glass Reactors has a Pg13.5 thread for screwing into the stainless steel head plates (1) of DASbox Bioreactors. • A silicone O-ring (3) is used to seal the construction. Regularly check the O-ring for cracks and damage and replace it if necessary. Screw the lip seal assembly into the middle Pg13.5 port in the head plate and tighten it. Next, attach impellers for cell culture or microbiology applications to the shaft as required. Loosen the set screw on the impeller using an Allen key until the impeller can be pushed along the stirrer shaft. Fix one or two impellers on the stirrer shaft depending on the application and requirements. The set screws are wear parts and must be replaced regularly. After mounting the impellers, close the bioreactor with the head plate. Note the following when attaching the motor to the lip seal assembly: • The latches (2) at the top end of of the lip seal assembly must engange the groove (1) in the motor drive ring. The latches (2) at the top end of the lip seal assembly must engage in the groove (1) in the motor drive ring. The motors for single-use bioreactors are connected by magnetic coupling to the bioreactor. You can find relevant information in the BioBLU 0.3c/f Instructions for Use. Hardware and operation DASbox® Mini Bioreactor System English (EN) 5.7 Cultivation – a typical workflow The following figure shows a typical process scheme for working with a parallel DASGIP Bioreactor System. It shows the sequence of individual work steps together with the approximate times required. Fig. 5-7: Work steps in a typical workflow for parallel DASGIP® Systems 73 74 Hardware and operation DASbox® Mini Bioreactor System English (EN) Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) 6 Software - DASGIP Control 6.1 Short overview 6.1.1 Login The program symbol of DASGIP Control is displayed on the desktop of the process computer. Double click on the program symbol to start DASGIP Control. Enter your user Name and the corresponding Password. Click Login to continue. 75 76 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) 6.1.2 Overview of Operations DASGIP Control is subdivided into three areas (see Fig. 6-1): Fig. 6-1: 1 2 Overview of DASGIP Control 3 Navigation area This area includes three different tabs: – Overview with Navigation Tree – Instructions – Navigation for a quick switching between the different views Working Area This area displays the Summary and Process View Event Monitor Displays all events, interventions and error messages Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) 6.1.3 New Workflow Click File in the menu and select New Workflow. Alternatively, click on the corresponding symbol in the task bar. Please refer to the DASGIP Control User Manual for more details. 6.1.4 Functional Overview DASGIP Control displays for the Navigation area three different views - Overview, Instructions, and Navigation - at the left side (refer to the DASGIP Control User Manual for complementary information). When you have started a process from the bioprocess controller, DASGIP Control displays the process name on the Overview tab under the branch Running in the Navigation Tree. 77 78 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Summary View Use the Navigation tab for a simple and intuitive operation within DASGIP Control. You can easily switch between Summary and Process View of different setups (vessels) of a process at any time. Click on the Xxx.Control button to display a Summary View, which gives a summary of all process values like pH, DO value(s), gas mixture, gassing rate and feeding volumes for a set of vessels selected to be operated in parallel (Fig. 6-2:). Large numerical display elements facilitate an easy reading of all relevant process values. The displayed views vary depending on your system. The following shows a Summary Views of DASGIP Control illustratively Fig. 6-2: Fig. 6-2: Summary View Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) The operating status of different sensors and actors (feeding pumps, gasmixer, impellers, etc.) is displayed by color coded symbols and monitored. The following table 6-1 shows the color definitions for the symbols. Tab. 6-1: Color definitions of Symbols Symbol Definition Green symbols indicate active elements. Orange symbols indicate elements in manual mode or operated by the bioprocess controller. Gray symbols indicate elements in standby mode or inactive elements. Blue symbols indicate elements in idle mode. Blue symbols indicate elements in error mode. To get the full access to each vessel in the subordinated Process View later on, you have to activate all sensor and control options in the Summary View superordinately, which means that all symbols are displayed in green. Additionally, you can click on each symbol to display the current process values of all affected reactors on the right side in the Summary View. The operating status of different sensors and actors (feeding pumps, gasmixing, impellers, etc.) is displayed by color coded symbols and monitored. The following table 6-2 shows the color definitions. 79 80 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Tab. 6-2: Color definitions of edit boxes Gray fields indicates read-only values. Values of white fields can be edited. Click on a white field to edit the value Gray fields with small triangles in the right upper corner indicate values, which should be analyzed before changing. Click on the triangle for editing. – Please note that this interrupts the automatic mode (gray) and switches into a manual one (white). Process View The Process View (Fig. 6-3:) displays all relevant process values to single setups / vessels. Switch to the Prozess View of a single setup by clicking on the corresponding Setup button on the Navigation tab. When you have started a process, the Process View displays all color coded transferred controller states. The following show a Process View of DASGIP Control illustratively. Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Fig. 6-3: Process View Please check and activate / deactivate each function according to your needs. Different symbols are available for vessels, sensors and controllers as well as actors like pump, gassing and agitation Controllers like pH and DO (i.e. DO Control) can operate actors like pumps, gas mixing or agitation. In this case orange symbols indicate actors which are controlled interdependently. The color indicates that direct access it is not possible. You have to inactivate the controller itself first by clicking on the white point of the symbol e.g. of DO Control to get access to the interdependent gas mixing. After a short delay the gas mixing symbol will turn from orange to gray and can be operated manually. Click on the vessel symbol to display the vessels parameters at the right side of the working area. 81 82 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Click on a controllers symbol to display the corresponding controller parameters. The parameter dialogs are subdivided in three tabs: • Details shows all relevant parameters at a glance. Edit boxes are displayed in white. • Profile (See “Define or Adapt Set-point Profiles” on page 83.) • Alarm (See “Alarm Option” on page 84.) Please refer to the DASGIP Control User Manual for complementary information. The following table 6-3 lists parameter definitions. Tab. 6-3: Parameter Definitions Abbreviation Parameter Definition State State The operation state of a controller is displayed as Busy, Manual, Inactive or Error. P Proportional Value The proportional value (P-Tune) directly determines the intensity of the control. The higher the chosen value, the more invasive is the control. Ti Reset time The reset time (Ti) influences the integrating part and therefore the sensitivity of the control. The larger the value, the smaller the effect on the control. The control becomes smoother, but slower at the same time. Td Derivation time Differentiator Min Lower limit Minimum value of the control variable (Anti-Reset-Windup). Max Upper limit Maximum value of the control variable (Anti-Reset-Windup). Deadband Deadband Control deadband for a two-sided control. Slope Slope Sensor slope calibration parameter. Offset Offset Axis shift as calibration parameter. Calibration Calibration Flow rate at 100 % output. Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Please note that not each 3rd party bioprocess controller supports all parameters. Therefore the Details tab in DASGIP Control displays differing parameters depending on the integrated controller. Define or Adapt Set-point Profiles DASGIP Control provides a very easy to use and flexible way to enter and edit controller profiles. Click on a controller symbol in Process View to open the corresponding values in Details at the right side. Click on the Profile tab to enter and edit the corresponding profile options. The drop down menu of Profile Source gives the opportunity to choose between different time or value based profiles (a detailed list of all profile sources is given in chapter 3.3.2 of the User Manual DASGIP Control). 83 84 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Each profile is sectioned into the three distinct presentation modes Type, Data and Graph: Type comprises three different profile types. Choose Constant Value, Datatable or Function as profile type. Use the Data tab to enter profile values and Graph to show the corresponding profiles graphically (please refer to chapter 3.3.2 and 3.7 of the User Manual DASGIP Control for more details). Alarm Option If the alarm option is activated, DASGIP Control displays alarm options on an Alarm tab within the specified sensor values separately. Click on the title bar or a process value (e.g. DO.PV) to display the corresponding sensor values. There is an additional Alarm tab, if the alarm option is activated. Click on the Alarm tab to adjust the alarm settings. Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Please note that all alarms are inactive until the inoculation clock (Inoculation) is started. Offline Values You can supplement the process documentation with offline values in DASGIP Control. These values determined by external analyzers can be entered together with the time stamp of determination directly or later on. They will be stored as an event in the database and can be displayed together with all other process values e.g. in a diagram. Click on the Offl. symbol in the Process View to display the dialog at the right side. You can enter up to four Offline values and user events by default. Additionally, you can specify the name as well as the engineering unit. Please refer to DASGIP Control User Manual chapter 5.1 for details. Chart Control Panel. During a process run all values are visualized in online charts in the Summary or Process View (Fig. 6-4:) at the bottom of the window. 85 86 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Fig. 6-4: Process View with Chart Control Panel Click on the corresponding curve button to hide or display a curve on the diagram. Click on the chart, hold the mouse button and drag a box to zoom in an area. Click on the x- or y-scale and drag down the appearing double line to display a legend with corresponding values. Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Click on the Tools button on the upper left edge of the Chart Control Panel to display further options. You can display all available data (Show all curves) or just data of determined time intervals (Zoom to last hour‘s data). You can configure and save chart settings individually (Save User chart settings). Please refer to the User Manual DASGIP Control chapter 6.1.1. for more details. Event Monitor The Event Monitor at the bottom of the DASGIP Control window displays all events, interventions and possible error messages of the connected bioprocess controllers during the process runtime. The following table 6-4 shows the color code of the Event Monitor. Tab. 6-4: Color code for the Event Monitor Color Description green message yellow warning red alarm 87 88 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Plant Overview The Plant Overview gives a complete tabular overview of the current process status and eases the access to main procedures’ process values. Additionally, the already logged process values of the running process can be viewed graphically and displayed according to individual points of view. Therefore you can combine any variables even historical ones for trend analysis. Click on the Navigation tab or on the plant name (XXX Plant) in the Navigation Tree to start the Plant Overview. Click on single values or select all values of a column by clicking on the column header. Your selection is marked by color and you can add values by pressing thekey and clicking simultaneously. Right click on the selection to display the context menu. Click History to display a diagram based on the selected values. Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Click in the diagram to display the integrated ruler, which shows the numeric values of the curve(s). Click and drag a frame in the diagram to zoom in or out. Create and print a diagram for documentation at any time. Click on the Tools button for the corresponding options. You can save and reload up to seven individually created diagram settings. 89 90 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) 6.2 Functions 6.2.1 Gassing When configuring cultivation processes, any profiles for gassing rates and gas compositions can be defined in advance. For the duration of the process, several options are available to intervene in the control and to change current process values: In the DASGIP Control software process view, click on the Gassing symbol to display the gassing control parameters. Change current set-points by clicking on the desired fields and overwriting the respective values contained there. Gassing as part of DO cascade control The cascade control for the oxygen concentration which is implemented in the software ensures that the parameters listed below are successively increased in order to maintain the predefined set-point for the O2 content of the culture: • Oxygen content in the inlet gas (XO2) • Gas flow (F) • Stirrer speed (N) The order of the parameters changed depends on the field of application. • With cascade control in microbiology applications, the gassing rate is increased as the last step as the possible formation of foam is a critical factor here. • In contrast, high shear forces are not desired in cell culture applications. Consequently, the stirrer speed is not considered, or optionally only increased as the last parameter, in the cascade for cell culture applications. Please refer to the User Manual DASGIP Control for detailed information on configuring gassing profiles, setting specific parameters, special gassing strategies and options for intervention during the process run. Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) 6.2.2 Temperature control During the configuration of cultivation processes, individual set-point profiles can be defined for the process temperature. During the process, several options are available to intervene in the temperature control and to change current values. In the DASGIP Control software process view, click on the symbol for temperature control Temperature to display the parameters for temperature control. Change current set-points by clicking on the desired fields and overwriting the respective values contained there. Please refer to the User Manual DASGIP Control for detailed information on configuring set-point profiles and setting control parameters. Options for intervening during the process run are also described here. 6.2.3 Agitation During the configuration of cultivation processes, any set-point profiles can be defined for agitation. During the process, several options are available to intervene in the agitation control and to adapt current values according to your requirements. In the DASGIP Control software process view, click on the symbol for agitation control Agitation to display the parameters for agitation control. Change current set-points by clicking on the desired fields and overwriting the respective values contained there. Agitation as part of DO cascade control The cascade control for the oxygen concentration which is implemented in the software ensures that the parameters listed below are successively increased in order to maintain the predefined set-point for the O2 content of the culture: • Oxygen content in the inlet gas (XO2) • Gas flow (F) • Stirrer speed (N) 91 92 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) The order of the parameters changed depends on the field of application. • With cascade control in microbiology applications, the gassing rate is increased as the last step as the possible formation of foam is a critical factor here. • In contrast, high shear forces are not desired in cell culture applications. Consequently, the stirrer speed is not considered, or optionally only increased as the last parameter, in the cascade for cell culture applications. Please refer to the User Manual DASGIP Control for detailed information on configuring set-point profiles and setting control parameters. Options for intervening during the process run are also described here. 6.2.4 Addition of media During the configuration of cultivation processes, you can define individual feeding profiles. Another option of adding substrate is available via DO based feeding. Here, the addition of substrate is controlled via the O2 content of the medium. This strategy allows for substrate-limited feeding. For the duration of the cultivation process, several options are available to intervene in the feeding control and to change current process values i ndividually. In the DASGIP Control software process view, click on the symbol for feeding Feeding to display the parameters for feeding control. Change current set-points by clicking on the desired fields and overwriting the respective values contained there. Please refer to the User Manual DASGIP Control for detailed information on configuring feeding profiles and special feeding strategies. It describes in detail the possibilities of intervention during the process run. Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) 6.2.5 Monitoring During the configuration of cultivation processes, you can configure individual set-point profiles for pH and DO. In addition, you can configure parameters for the level and anti foam option or the redox measurement. For the duration of the cultivation process, you can actively intervene in the control and change current values. In the DASGIP Control software process view, click on the symbol for the desired control (pH, DO, etc.) Control to display the parameters for the respective control. Change current set-points by clicking on the desired fields and overwriting the respective values contained there. Please refer to the User Manual DASGIP Control for detailed information on configuring set-point profiles and setting control parameters. It also contains details on configuring the options level control with addition of anti foam and redox measurement. 93 94 Software - DASGIP Control DASbox® Mini Bioreactor System English (EN) Maintenance DASbox® Mini Bioreactor System English (EN) 7 Maintenance 7.1 Cleaning and decontamination 7.1.1 Surfaces, modules Simply wipe off surface contamination with a damp, lint-free cloth. If necessary, use a mild cleaning agent or disinfectant (such as 80% EtOH). Or clean the surfaces according to the standard SOP in your laboratory. NOTICE! Risk of damage to device from corrosive media. Prolonged contact with aggressive media can attack surfaces. Remove any acid and base residues by thoroughly washing with water as soon as possible. CAUTION! Risk of damage to device from improper use of cleaning agents. Do not use any aggressive, corrosive or abrasive disinfectants or cleaning agents. If you have any questions about the compatibility of cleaning agents and disinfectants or their ingredients, please contact a service technician at the Eppendorf Bioprocess Center. WARNING! The DASbox devices are not splash-proof. 7.1.2 Cleaning feed tubing and pump tubing Media components which crystallize in the DASbox Feed Tubing can lead to irreversible clogging of the supply lines. Media residues remaining in the supply lines may also encourage the growth of microorganisms in the supply lines. After each use, thoroughly rinse the feed tubing and pump tubing with water and/or ethanol (do not use ethanol for C-Flex® supply lines). Cleaning procedures for feed tubing are implemented in the DASGIP Control software. They can be adapted to suit your requirements at any time. Please refer to the User Manual DASGIP Control. 95 96 Maintenance DASbox® Mini Bioreactor System English (EN) NOTICE! Limited chemical resistance of C-Flex® tubing! C-Flex® tubing is not suitable for use with alcohols and organic solvents. Do not rinse feed tubing made of C-Flex® with ethanol for cleaning purposes. CAUTION! Danger from squirting of liquids! The blockage of feed tubing may lead to a pressure build-up in the system and the squirting of liquids! When working with the DASbox, wear personal protective equipment, especially goggles! 7.1.3 Decontamination before service or shipment Before shipping DASGIP Systems or system components for service purposes or before an on-site service, make sure to observe the information in our document "Return delivery information". It contains a decontamination certificate ("Cleaning and Decontamination Form") which must be completed in advance and sent to DASGIP. Download the document "Return delivery information" from our homepage: (http://www.dasgip.com), see Service/Technical service "Return delivery information"). WARNING! Risk to health from contaminated devices! Observe the notes on the decontamination certificate. It is available as a fillable PDF document on our website (http://www.dasgip.com), see Service/Technical service "Return delivery information"). Send a copy of the completed decontamination certificate to DASGIP in advance and attach the original on the outside of the packaging. If you have any questions, contact our Service Department (bioprocess-support@eppendorf.de). Maintenance DASbox® Mini Bioreactor System English (EN) 7.2 Service DASbox System components are thoroughly checked and tested before delivery. To ensure continuous error-free operation, we recommend that the system is serviced annually by a service technician from the Eppendorf Bioprocess Center. Information about the support services is available on our website www.dasgip.com. If you have any questions or problems concerning the operation of DASbox, contact our Service Department: bioprocess-support@eppendorf.de +49(0)2461 980-0 7.3 Additional information on care and maintenance 7.3.1 DO sensors (Clark type) NOTICE! Risk of sensor damage from improper handling. ALWAYS observe the information and instructions provided by the sensor manufacturer with priority when handling, inspecting and cleaning your DO sensors. Sensor inspection and replacement of electrolyte solution Depending on how the DO sensors are used and for how long, the electrolyte solution must be replaced at regular intervals, at least after 6 months. The following section describes how to replace the electrolyte solution for the Mettler Toledo sensors which are used as standard. CAUTION! The O2 electrolyte solution is strongly alkaline! Wear suitable protective clothing, especially gloves, as liquid will inevitably come into contact with your hands during the process of assembling the membrane body. Avoid contact with skin and mucous membranes, especially the eyes. Rinse contaminated areas with plenty of water. Wear protective goggles! 97 98 Maintenance DASbox® Mini Bioreactor System English (EN) 1. Unscrew the cap from the sensor body. 2. Carefully remove the cap from the sensor. 3. Remove the membrane body from the cap. If the body is too firmly connected, gently push it out with your fingertip. 4. Rinse the inner body with demineralized water and gently pat it dry with paper towels. 5. Visually inspect the O-rings and the trapeze rubber for mechanical defects. 6. Half fill the membrane body with O2 electrolyte solution. 7. Remove air bubbles by gently tapping on the membrane body. 8. Hold the sensor in a vertical position and slide the body membrane onto the inner body. 9. Remove excess electrolyte solution with a paper towel. 10. Carefully slip the cap over the fitted membrane body and tighten it firmly. Note: After each replacement of electrolyte solution, a calibration must be performed to determine the zero point and slope of the sensor. After each replacement of electrolyte solution, the sensor has to be completely polarized (2 - 6 h) and a calibration carried out afterwards! Also observe the manufacturer's instructions in the relevant documentation for the DO sensors. 7.3.2 pH sensors NOTICE! Risk of sensor damage from improper handling. ALWAYS observe the information and instructions provided by the sensor manufacturer with priority when handling, inspecting and cleaning your pH sensors. Regeneration of pH sensors The functioning of pH sensors that appear visually intact but are sluggish in responding can often be significantly improved by a special cleaning process. Soak pH sensors which have been stored in dry conditions or in water for an extended period in 3 mol/l KCl solution for several hours. Maintenance DASbox® Mini Bioreactor System English (EN) The method of cleaning varies depending on the type of deposits visible: Inorganic deposits Store the sensors in 0.1 mol/l HCl or 0.1 mol/l NaOH for several minutes. Oily or organic deposits Rinse the sensors with ethanol or acetone. Proteins Store the sensors in pepsin/HCl solution for several hours. Sulfide contamination (black diaphragm) Store the sensor in Thiourea/HCl solution until the discoloration of the diaphragm is removed. 7.3.3 MP8-pHpO module Regularly replace pump head tubing The pump head tubing that is inserted into the pump tube guide is a wear part. If possible check for clearly visible signs of wear before each long experiment. Replace tubing with strong black abrasion, kinking or very irregular wall thickness. Another sign of wear is if the calibration value strongly deviates from the original value. Note: Make sure to replace tubing in time to avoid leaks and to maintain high feeding accuracy. Regularly replace the back-up batteries The back-up batteries of the MP8-PHPO module guarantee the storage of internal data whenever the module is switched off. To avoid a loss of internal data, the batteries have to be renewed regularly. NOTICE! Risk of internal data loss and equipment damage. Replace the batteries as soon as the battery replacement message is displayed in the DASGIP control software event log. Ensure that the MP8-PHPO module is switched on while replacing the batteries. Replacing the batteries while the device ist switched off will lead to data loss and equipment damage. 99 100 Maintenance DASbox® Mini Bioreactor System English (EN) The holders for the back-up batteries are located in the lower part of the MP8-PHPO module in the storage compartment for feed tubing. Obey the following instruction to replace the batteries. 1. Ensure that the MP8-PHPO module is switched on. The status LED of the device has to be illuminated green. 2. Open the flap of the lower storage compartment. The holders for the back-up batteries are located on the right and left side in the storage compartment. 3. Unscrew the lid of the battery holder by a quarter turn in a counterclockwise direction. Use a flat head screwdriver if necessary. 4. Now remove all 3 batteries from the holder. The batteries are magnetic. They can easily brought out using a magnetic stick (e.g. a stir bar retriever). Maintenance DASbox® Mini Bioreactor System English (EN) 5. You will find 3 Batteries per holder. Always replace all 6 back-up batteries of the module. Only use approved batteries (see chapter 10.3). 6. When replacing the batteries, the positive terminal of the batteries must show to the outside. 7. Check the orientation of the battery compartment lid. Metal flaps and slot on the lid must be in horizontal orientation to close the lid. 8. Align the lid of the battery holder and close it by turning the lid clockwise until safely locked. 101 102 Transport, storage and disposal DASbox® Mini Bioreactor System English (EN) 8 Transport, storage and disposal 8.1 Transport When sending in products for service purposes as well as during transport to another installation location, ensure proper packaging is used. CAUTION! Risk of personal injury and material damage when lifting and carrying heavy loads. The DASbox is heavy. Lifting and carrying the device can lead to back injuries. The hooks at the front and the grip at the back of the DASbox are not handles. They are only designed for moving the DASbox on telescopic rails or for lifting the lid during service work. NEVER use the hooks and the grip on the DASbox as handles. The DASbox must be transported by least two people. Lift the DASbox by reaching under the device. Use a transport aid (such as a dolly) to transport the device over longer distances. NOTICE! Risk of transport damage! Use the product's original packaging for transport and return of products. Please also observe the instructions in "How to pack…" in the document "Return delivery information". Download the document from our website (http://www.dasgip.com), see Service/Technical service "Return delivery information"). or contact our Service Department (bioprocess-support@eppendorf.de). If the original packaging of the product is no longer available, the customer is responsible for providing proper and suitable packaging. Original packaging can be requested from DASGIP at any time. 8.2 Storage Observe the ambient conditions during storage. Ambient temperature during storage 0 - 50°C Max. relative humidity during storage 90% rH, non-condensing. Transport, storage and disposal DASbox® Mini Bioreactor System English (EN) 8.3 Disposal If disposing of products or product components, make sure to observe the relevant legal regulations. As the disposal regulations may differ from country to country in the European Union, please contact us if necessary (bioprocess-support@eppendorf.de). Environmentally sound disposal of electrical and electronic equipment in compliance with EU Directive 2002/96/EC In order to comply with EU Directive 2002/96/EC, please observe the applicable national regulations when disposing of DASGIP devices. *For Germany the law regarding waste electrical and electronic devices (ElektoG) applies: Under this Directive, no old devices may be disposed of via the public municipal waste/collection system. For DASGIP devices located in Germany you can return such devices to us for scrapping. Decontaminate all the parts you would like to dispatch for disposal. Observe the following safety instructions and the chapter 7.1. WARNING! Risk to health from contaminated devices! Observe the notes on the decontamination certificate. It is available as a fillable PDF document on our website (http://www.dasgip.com), see Service/Technical service "Return delivery information". Send a copy of the completed decontamination certificate to DASGIP in advance and attach the original on the outside of the packaging. If you have any questions, contact our Service Department (bioprocess-support@eppendorf.de). 103 104 Transport, storage and disposal DASbox® Mini Bioreactor System English (EN) 8.4 General return delivery information When returning products or product components, e.g., for servicing or disposal, and also before an on-site service, make sure to observe our general return delivery information. Download the document "Return delivery information" from our website. (http://www.dasgip.com), see Service/Technical service "Return delivery information". The document "Return delivery information" as a fillable PDF document contains the following forms: • Decontamination certificate ("Cleaning and Decontamination Form") • Service form ("Service and Repair Form") • Packaging instructions for DASGIP Modules ("How to pack...") WARNING! Risk to health from contaminated devices! Observe the notes on the decontamination certificate. It is available as a fillable PDF document on our website (http://www.dasgip.com), see Service/Technical service "Return delivery information". Send a copy of the completed decontamination certificate to DASGIP in advance and attach the original on the outside of the packaging. If you have any questions, contact our service department (bioprocess-support@eppendorf.com). Technical data DASbox® Mini Bioreactor System English (EN) 9 Technical data 9.1 Ambient conditions Ambience For indoor use only Ambient temperature during operation 5 – 40°C (41 – 104°F) Max. relative humidity during opera- 80% rH, non-condensing tion Maximum operating altitude 6000 ft above sea level Ambient temperature during storage 0 - 50°C Max. relative humidity during storage 90% rH, non-condensing WARNING! The DASbox System must not be used outdoors. 9.2 DASbox® System 9.2.1 Power supply Voltage (DASbox Base Unit and MP8-PHPO module) 100-240 VAC Operating frequency (DASbox Base Unit and MP8-PHPO module) 50/60 Hz Typical power consumption (DASbox Base Unit) Approx. 510 W Typical power consumption (MP8-PHPO module) Approx. 50 W Fuses (DASbox Base Unit) 2 x 10 AT, 250 V Fuses (MP8-pHpO multi pump and sensor module) 2 x 2.5 AT, 250 V Protection class I Protection degree (IP Code) IP21 (Ingress Protection Rating) Overvoltage class II Back-up battery Varta High Energy 4906 1.5 V, AA, Alkaline 105 106 Technical data DASbox® Mini Bioreactor System English (EN) 9.2.2 Weight / dimensions Dimensions W/H/D DASbox System (four-fold unit w/o autoclavable carrier) 30/49/62* cm incl. DASbox Base, MP8-pHpO w/ Storage (11.8/19.3/24.4* in) DASbox System (four-fold unit w/o autoclavable carrier) 30/66/62* cm incl. DASbox Base, MP8-pHpO w/ Storage and one additional DASGIP module (11.8/26/24.4* in) DASbox Base 30/21/62* cm (11.8/8.3/24.4* in) MP8-pHpO w/ Storage 30/29/33* cm (11.8/11.4/13* in) DASGIP module 30/20/33* cm (11.8/7.9/13* in) Autoclavable carrier 30/7/14 cm (11.8/2.6/5.5 in) *Note at least additional 10 – 15 cm (3.9 – 5.9 in) depth for cable and heat dissipation Weight DASbox Base Unit w/o accessories approx. 28.6 kg MP8-pHpO w/ Storage approx. 9 kg Standard DASbox System (four-fold unit) approx. 44 kg 9.2.3 Bioreactors Reactors Glass reactors / single-use vessels Sterilization Autoclavable / presterilized Cell culture working volume Glass: 60 – 250 ml / single-use: 100 – 250 ml Microbiology working volume Glass: 60 – 250 ml / single-use: 65 – 250 ml Total volume Glass: 350 ml / single-use: 380 ml Dimensions (ID, H) 64 mm/120 mm Technical data DASbox® Mini Bioreactor System English (EN) 9.2.4 Agitation Drive Overhead drives Glass: directly coupled / single-use: magnetically coupled Speed ranges Glass: 20 – 2000 rpm / single-use: 20 – 2000 rpm Cell culture impellers Glass: marine type / single-use: pitched blade Microbiology impellers Rushton type 9.2.5 Gassing Gassing TMFC; overlay and/or submerged Gas mixture (standard) N2, air, O2 and/or CO2 Flow control & mixing 16 independent mass flow controllers Flow ranges Cell culture: 0-5 sl/h / microbiology: 0-25 sl/h Off-gas condensation Liquid free using Peltier elements Off-gas analysis Optional; with additional module DASGIP GA4 9.2.6 Dispensing Feeding lines per reactor 2 (optional 4) Feeding rates (depending on tubing diameter) 0.3 – 9.5 ml/h up to 13 – 420 ml/h 9.2.7 Monitoring and control Temperature control Liquid-free heating and cooling (Peltier elements) Temperature range 10 – 60°C at 25°C RT Temperature sensor Accuracy class A RTD pH control Cell culture: CO2/base Microbiology: Acid and/or base other configurations possible in each case 107 108 Technical data DASbox® Mini Bioreactor System English (EN) DO control Cell culture: Cascade (O2 concentration, gassing rate) Microbiology: Cascade (stirrer speed, O2 concentration, gassing rate) other configurations possible in each case Redox measurement (ORP) optional; as an alternative to level control Level/ anti foam control Optional; as an alternative to Redox measurement Optical density (OD) Optional; with additional module DASGIP OD4 Technical data DASbox® Mini Bioreactor System English (EN) 9.3 Ports - specifications Mains connections Mains connection DASbox Base Unit (all versions): 100-240 VAC; 50/60 Hz MP8-pHpO Module (all versions): 100-240 VAC; 50/60 Hz Microfuses Dimensions 5x20 mm; rated voltage at least 250 V, characteristic slow-blow; nominal curent 2.5 A (MP8-pHpO); 10 A (DASbox Base Unit) Communication "AUX"; "DTP out" SUB D9M / RS232 plug "DTP in" SUB D9F / RS232 connector DASbox Base Unit USB port 5 V / 100 mA; 5 V +/- 0.25 V internal power supply Stas 200 type ports; Input limits: Max. 2.5 A for each port (depending on the incoming power supply 100-240 VAC, 50/60 Hz at the mains connection unit for the external power supply). Pt 100 (temperature sensors) Input limits: 90 – 160 Ohm Measuring range: -25°C – 116°C (± 1 °C) Condenser (Off-gas condensers) Typically 10 W per condenser Gas IN; DASbox Base rear side: 6 mm push-in connectors Gas OUT Next to the wells for bioreactors: 4 mm push-in connectors External I/O 0/4 – 20 mA or 0 – 10 V inputs and outputs *optional: external interfaces MP8-pHpO Module pH Sensors Input limits: - -472 mV – 472 mV Measuring range: pH 0 – pH 14 (± 0.01 pH) (at a sensor temperature of max. 67°C) DO Sensors DO Measurement: Input limits: 0 – 400 nA (± 0,5 nA) Measuring range: 0 – 500 % DO Integrated temperature measurement (NTC): Input limits: 1.7 – 10000 kOhm Measuring range: -10°C – 140°C (± 0.2 K) Redox Sensors Conductivity measurement Input limits: 0 – 20 mS Redox potential measurement Input limits: -2000 – 2000 mV (± 1 mV) 109 110 Technical data DASbox® Mini Bioreactor System English (EN) 9.4 Connection assignments 9.4.1 Communication ports (DTP in/out, AUX) The RXD, TXD, GND, RTS, CTS supply lines of the DTP in and DTP out ports are directly connected to ‐one‐ another. Pin DTP in DTP out 1 AUX Alarm 2 TXD TXD RXD 3 RXD RXD TXD GND GND GND 4 5 6 Alarm (normally closed) 7 CTS CTS 8 RTS RTS 9 Alarm (normally open) In a fault-free state, the pins 1 and 6 are connected to each other (normally closed). In the event of an error, the pins 1 and 9 are connected to each other (normally open). Only devices which meet the requirements of IEC 950/EN 60950-1 (UL 1950) standards may be connected to the interfaces. CAUTION! The maximum load capacity of the relay contacts is 30 VDC / 1 A. 9.4.2 RS485 (optional) The RS485 communication ports are galvanically insulated from the rest of the device electronics. The pin assignment complies with the specification "Modbus Over Serial Line Specification and Implementation Guide Version 1.0". The B/D1, A/DO and Common supply lines of the two RJ45 connectors are directly connected to one another. Technical data DASbox® Mini Bioreactor System English (EN) Pin 1-4 Pin 5-8 Description 1 5 A/D0 2 6 3 7 + 5 V auxiliary power 8 Common 4 9.4.3 Description B/D1 Sensor connectors pH connector Pin Assignment 1 pH+ 2 3 GND Pin Assignment 1 NTC 2 UVisiFerm (10 V) 3 GND DO connector 4 5 DO 6 Upol 111 112 Technical data DASbox® Mini Bioreactor System English (EN) Level connector Pin Assignment 1 LVL+ 2 GND Redox connector (optional, instead of level) Pin Assignment 1 RDX 2 3 GND Pin Assignment 1 Pt 100 port A Pt connector 2 3 Pt 100 port B 4 Pt 100 port B´ To connect the Pt100-type temperature sensors, Binder 680-type 4-pin connectors are used. To ensure the proper functioning of the three wire circuit, the B and B’ signals have to be directly interconnected at the sensor element of the temperature sensor. Note: The measuring amplifiers of the primary and secondary temperature sensors are galvanically insulated from the rest of the device electronics. The measuring amplifiers have a common reference potential. Ordering information DASbox® Mini Bioreactor System English (EN) 10 Ordering information 10.1 DASbox base unit Order no. (international) Description 78525185 DASbox Overhead Drive for glass reactors (20-2000 rpm), incl. cable L 1.6 m 78525186 DASbox Overhead Drive for Single-Use Vessels (20-2000 rpm), incl. cable L 1.6 m 78201321 Actuator unit for DASbox peltier off gas condenser, incl. cable L 1.6 m Off-gas condenser adapter incl. insulation 78201323 for DASbox glass reactor 78201322 for DASbox Single-Use Vessel 78109111 Insulation for Off-gas condenser adapter 78590033 Exhaust Gas Condenser Controller EGC4, 4 ports for Peltier Condensers 78109112 DASbox autoclavable carrier for 4 vessels, stainless steel, labeled 78109113 DASbox autoclavable carrier for media bottles, stainless steel, labeled 78109030 Holder for DASbox autoclavable carrier 78201035 Flask 100 mL with neck GL45, transparent, with cap 78201078 Flask 500 mL with neck GL45, transparent, with cap 78510311 DASGIP head gear for reservoir bottles with GL45 neck, silicone, female luer 78903224 GL45 screw cap, PBT, with port D 34 mm 78201311 Pull-out system for one DASbox base unit 78201146 Sealing ring for one well of DASbox base unit, silicone 78105217 Gas tubing PA, OD 4 mm/ID 2,5 mm, outside tolerated, oxygen resistant, uncolored 78105218 Gas tubing PA, OD 6 mm/ID 4 mm, outside tolerated, oxygen resistant, uncolored 113 114 Ordering information DASbox® Mini Bioreactor System English (EN) Order no. (international) Description 78704128 Disassembly tool for Push-In OD 4 mm and OD 6 mm, plastic 10.2 System / cables Order no. (international) Description 78702579 Cable converting Hirschmann STAS200 plug to IEC 320 C14 connector, L 60 cm 78702468 L 43 cm 78535021 Terminator connector for DTP network version 2 78100242 DTP control cable, L1m 78702562 DASGIP cable, 4pinM5, L 3 m, (for LCD monitor) Ordering information DASbox® Mini Bioreactor System English (EN) 10.3 MP8-pH4pO4 multi pump and sensor module Order no. (international) Description 78530260 MultiPump & Sensor Module MP8-PH4PO4LS w/ Storage, 8 pumps and 4xpH, 4xDO and 4xLeveL 78530261 MultiPump & Sensor Module MP8-PH4PO4RD4S w/ Storage, 8 pumps and 4xpH, 4xDO and 4xRedox 78705383 Back-up battery for DASGIP Module MP8-PHPO (Varta High Energy 4906 1.5 V, AA, Alkaline 78103314 Platinum RTD temperature sensor, 100 Ohm class A, OD 1.6 mm, L 150 mm, cable L 1.8 m 78522020 DASGIP pH/Redox cable with AK9 plug; L 3 m 78103207 pH Sensor MTI D 12 mm, L 120 mm, autoclavable 78103224 Redox Sensor MTI D 12 mm, L 120 mm, autoclavable 78522040 DASGIP DO cable with plug Type 82, L3m 78108016 Dust cap for DO Sensor with 4-pin connector (T82) 78108046 DO Sensor for Single Use Vessels DO sensor DASGIP D 4.7 mm, L 162 mm, incl. cable L 3m and calibration cap 78108018 DO Sensor MTI D 12 mm, L 120 mm autoclavable 78108003 DO-Membrane Kit MTI (4 membrane bodies, spare O-rings, 25 mL electrolyte) 78522031 DASGIP Level Sensor cable, L3m 78103117 DASGIP Level Sensor OD 4 mm, L 205 mm, L i165 mm Pump head tubing for MP8 pump, 78510119 Bioprene, ID 0.25/W 0.85 mm, female/female 78510118 Bioprene, ID 0.5/W 1.05 mm, female/female 78510236 Bioprene, ID 1.0/W 1.05 mm, female/female Feed line C-Flex, 78510309 ID 0.8 mm, L 1 m, w/ 2x Luer lock fittings, male/male 78510310 ID 0.8 mm, L 2 m, w/ 2x Luer lock fittings, male/male 115 116 Ordering information DASbox® Mini Bioreactor System English (EN) Order no. (international) Description Feed line PTFE, 78510242 ID 0.8 mm, L 2 m, uncolored w/ 2x Luer lock fittings, male/male 78510241 ID 0.8 mm, L 1 m, uncolored w/ 2x Luer lock fittings, male/male 78510240 ID 0.8 mm, L 0.5 m,uncolored w/ 2x Luer lock fittings, male/male Feed line silicone, 78510246 ID 0.8 mm, L 2 m, w/ 2x Luer lock fittings, male/male, Pumpsil 78510245 ID 0.8 mm, L 1 m, w/ 2x Luer lock fittings, male/male, Pumpsil Ordering information DASbox® Mini Bioreactor System English (EN) 10.4 DASbox Bioreactors 10.4.1 Single-Use Vessels Order no. (international) Description 78903508 BioBLU 0.3c single-use vessel, cellculture, 4 pack, pre-sterilized, blue 78903509 BioBLU 0.3f single-use vessel, microbial, 4 pack, pre-sterilized, blue 10.4.2 Glass Reactors Order no. (international) Description 76DS0250ODSS DASbox Vessel Type DS0250ODSS for cell culture applications (Pitched blade impeller, 60mL - 250mL, overhead drive) 76SR0250ODLS DASbox Vessel Type SR0250ODLS for microbiology applications (2x Rushton impeller, 60 mL - 250mL, overhead drive) 78903227 DASGIP GLS-80 vessel OD 70 x H 123 mm, flat bottom, with Eppendorf Logo 78107301 Stainless steel Headplate OD 77 w/ 6x Pg13.5, 4x D 4 mm dip tubes, 1x thermowell 78903226 Screw cap for OD 80 mm (GLS80) neck with hole ID 73 mm, PBT, blue 78201138 O-Ring 68 x 3 mm (IDxd) VMQ Si820 78107304 6 blade Rushton turbine OD 30 mm, ID 5 mm, stainless steel, for microbiology 78107325 3 blade impeller 30° pitch, OD 30 mm, ID 5 mm, stainless steel, for cell culture 78107326 Stainless steel pipe OD 4 mm/ID 2 mm, L 180 mm, with barb 78200119 Tubing clamp (Mohr) L 50 mm, nickel plated brass 78532034 Filter assembly type "Gas In" 78532035 Filter assembly type "Gas Out" 117 118 Ordering information DASbox® Mini Bioreactor System English (EN) Order no. (international) Description 78532036 Liquid feed connector assembly 78510145 DASGIP sampling accessory w/ swabable valve for vessels type BS/DS/SR/DR 78200077 Sampling valve male luer lock and swabable female luer lock, autoclavable 78200087 Cap for sampling valve male luer lock, autoclavable 78704149 Hex wrench w/ ball shaped head, size 1.5mm 78532282 DASGIP compression fitting ID 4 mm with PG13.5 male thread, all parts included: 78706417 O-Ring 4 x 1.5 (IDxd), Si820, red 78706351 Front ferrule PFA, for OD 4 mm DASGIP compression fitting 78706352 Back ferrule PFA, for OD 4 mm DASGIP compression fitting 78706406 O-Ring 14 x 2 (IDxd), VMQ70/Si820, red 78532283 DASGIP compression fitting ID 6 mm with PG13.5 male thread, all parts included: 78706416 O-Ring 6 x 1.5 (IDxd), Si820, red 78706353 Front ferrule PFA, for OD 6 mm DASGIP compression fitting 78706354 Back ferrule PFA, for OD 6 mm DASGIP compression fitting 78706406 O-Ring 14 x 2 (IDxd), VMQ70/Si820, red 78532284 DASGIP compression fitting ID12 mm with PG13.5 male thread, all parts included: 78706419 O-Ring 12 x 1.5 (IDxd), Si820, red 78706355 Front ferrule PFA, for OD 12 mm DASGIP compression fitting 78706356 Back ferrule PFA, for OD 12 mm DASGIP compression fitting 78706406 O-Ring 14 x 2 (IDxd), VMQ70/Si820, red Ordering information DASbox® Mini Bioreactor System English (EN) Order no. (international) Description 77102018 DASGIP triple port w/ 3x OD4mm tubing, w/M18x1.5 male thread, all parts included 78706415 O-Ring 4 x 1 (IDxd), VMQ70/Si973, red 78706406 O-Ring 14 x 2 (IDxd), VMQ70/Si820, red 77102006 Septumholder Pg13.5 male with septum ID 12 mm, stainless steel 78706409 Septumholder DASGIP fitting w/ Pg13.5 male thread, ID 12 mm port, stainless steel 78106309 Silicone rubber septa Pg13.5, OD 18 mm/ ID 12 mm 78525123 DASGIP Lipseal stirrer assembly Pg13.5 seat, stirrer shaft d 5 mm x Li 112 mm 119 120 Ordering information DASbox® Mini Bioreactor System English (EN) 121 Index Calibration procedures ........................................ 15 Carrier System (autoclavable) .............................. 67 A Abbreviations ......................................................... 7 Addition of media .................................................92 Addition of media - see Feed Agitation ...............................................................43 Speed limits ....................................................43 Technical data ...............................................107 Agitation control ...................................................91 Alarm Settings ...........................................................84 Alarm Option - DASGIP Control ...........................84 Alarms ..................................................................67 Cascade control ............................................. 90, 92 C-Flex® tubing .............................................. 54, 96 CIP - seeCleaning-in-place Cleaning Feed Tubing .................................................... 95 pH Sensors (Regeneration) ............................ 98 Pump tubing ................................................... 95 Surfaces .......................................................... 95 Cleaning agents ................................................... 95 Cleaning-in-place (CIP) .................................... 9, 54 Cleaning-in-place procedures .............................. 15 Ambient conditions ..............................27, 105, 106 CO2 Flowrates .......................................................... 5 Assembly Glass Reactor ..................................................71 Overhead Stirrer .............................................72 Colors Software Event Monitor .................................. 87 Software Symbols ........................................... 79 Autoclavable Carrier System ................................67 COM ports ............................................................ 33 Communication ............................................ 109 Install .............................................................. 36 Ports ............................................................... 29 Autoclave Feed tubing .....................................................55 pH-Sensors ......................................................58 AUX ......................................................................29 B back-up batteries ..................................................99 BioBLU® 0.3c/f Single-Use Vessel ......................13 Bioreactor Assembly .........................................................71 Bioreactors .....................................................13, 69 Description ......................................................13 Technical data ...............................................106 C Calibrate DO Sensors .....................................................59 pH Sensors ......................................................58 Pump head tubing ...........................................54 Sensors ............................................................58 Calibration factor FCal .........................................50 Condensate Frozen ............................................................. 48 Condenser adapter .............................................. 44 Condensers Component parts ............................................ 44 Connecting sockets ........................................ 30 Conductivity Sensors Connecting sockets ........................................ 33 Connect DASbox Base Unit .......................................... 35 Gas supply ...................................................... 40 Off-gas condensers ......................................... 38 Overhead drives ............................................. 38 Sensors ........................................................... 39 Temperature sensors - Base Unit ................... 38 Connection assignments .................................... 110 Control - see DASGIP Control 122 Cooling System .....................................................43 Disassembly tool .................................................. 40 Cultivation ............................................................73 Dispensing Continuous ..................................................... 51 Pulsed ............................................................. 51 Technical data .............................................. 107 D Danger Levels ................................................................6 Symbols .............................................................6 DASbox Base Unit - see DASbox Base Unit Bioreactors - see Bioreactors Carrier System ................................................67 Connect ...........................................................34 Features ..........................................................14 Not connected .................................................65 Storage ................................................11, 12, 65 System - general view .....................................11 System - rear view ..........................................12 Versions ............................................................5 DASbox Base Unit ..........................................11, 12 Connect ...........................................................35 Connecting sockets .........................................30 Functions ........................................................41 Rear .................................................................30 DASGIP Control ............................................................15 Service ..................................................................97 DASGIP Control Alarm Option ...................................................84 Chart Control Panel .........................................85 Event Monitor .................................................87 Login ...............................................................75 Process View ...................................................80 Summary View ................................................78 DASGIP Software ..................................................15 DASware ...............................................................15 Decontamination ............................................95, 96 Definition Of Set-point Profiles ........................................83 Software Parameters .......................................82 Definitions ..............................................................7 Deice Function .....................................................48 Display ................................................................. 65 Disposal ..................................................... 102, 103 DO Cable ............................................................... 39 Sensor for Single Use vessels ........................ 39 Sensor membrane .................................... 61, 62 DO cascade control .............................................. 91 DO connector ..................................................... 111 DO Sensors Calibrate ......................................................... 59 Connecting sockets ........................................ 33 For Single-Use Bioreactors ............................ 63 Handling instructions ..................................... 61 Maintenance ................................................... 97 Polarize ..................................................... 62, 63 Raw signal ...................................................... 62 Replacement of Electrolyte ............................ 97 Documents Other applicable ......................................... 5, 18 DTP cable ............................................................. 36 DTP in .................................................................. 29 DTP network ........................................................ 36 DTP out ................................................................ 29 E Edit boxes ............................................................ 80 Electrical connections requirements .................................................. 28 Electrical supply Install .............................................................. 35 Electrolyte solution DO Sensors ..................................................... 97 Errors and Alarms ................................................ 67 Delivery package ..................................................14 Event Monitor ................................................ 76, 87 Colors ............................................................. 87 Dimensions .........................................................106 Exhaust filter blockages ....................................... 47 123 Exhaust gas line insulation ............................44, 45 H External power supply ..........................................35 Head plate Typical Assignments ....................................... 70 F FCal ......................................................................50 Features DASbox ..................................................14 Feed Pump head tubing ...........................................49 Rates / Ranges ..........................................49, 50 Tubing .............................................................49 Feed tubing Autoclave ........................................................55 Cleaning ..........................................................95 Sterilize ...........................................................54 Feeding Continuous ................................................51, 56 Pulsed .............................................................51 Short ................................................................56 Filter blockages ....................................................47 I Inlet pressure Gases .............................................................. 21 Installation instructions ....................................... 27 Intended use ........................................................ 17 Warnings ........................................................ 18 Internal power supply .......................................... 35 L LCD Monitor ........................................................ 65 Connect .......................................................... 37 Port ........................................................... 30, 33 Level Cable ............................................................... 39 Connector ..................................................... 112 Sensors ........................................................... 59 Freezing of condensate ........................................48 Fuses ....................................................................29 Location Requirements ................................................. 27 G L-Sparger ............................................................. 70 Gas IN - see Gas inlets M Gas inlets ..............................................................31 Main illustration ................................................... 11 Gas Mixing System ...............................................42 Splitter Function .............................................42 Variants ...........................................................42 Mains connection .............................................. 109 Gas outlets ............................................................31 Mains disconnect switch ..................................... 29 Gas quality ............................................................28 Maintenance ........................................................ 97 Gas supply Connect ...........................................................40 Ports ................................................................31 Media addition - see Feed Mains connection unit ......................................... 29 Microfuses ......................................................... 109 Monitor - seeLCD Monitor Gassing .................................................................42 Technical data ...............................................107 Gassing tubes Loosen .............................................................40 Glass Reactor ........................................................13 Assembly .........................................................71 Glossary ..................................................................9 Monitor cable ....................................................... 37 Monitoring ..................................................... 57, 93 MP8-pHpO Module Connect .......................................................... 39 Function ......................................................... 49 Ports ............................................................... 33 Rear ................................................................ 33 124 Multi Pump and Sensor Module ..........................33 Peltier elements ............................................. 44, 48 MX4/4-3 Gas Mixing Module - see Gas Mixing System pH cable ............................................................... 39 N Navigation Area ....................................................76 New Workflow ......................................................77 Not connected ......................................................65 O Off-gas condensers ...............................................44 Adapter ...........................................................44 Assembly for glass reactors ............................45 Assembly for single-use vessels ......................46 Connecting sockets .........................................30 Deice ...............................................................48 Heating mode ..................................................48 Operate ............................................................47 Offline Values .......................................................85 pH connector ..................................................... 111 pH Sensors Autoclave ........................................................ 58 Calibrate ......................................................... 58 Connecting sockets ........................................ 33 Handling instructions ..................................... 60 Raw signal ...................................................... 61 pH sensors Regeneration .................................................. 98 Plant Overview ..................................................... 88 History ............................................................ 88 pO2 Sensors - see DO Sensors Polarize .......................................................... 62, 63 Ports Specifications ............................................... 109 Operating pressure Gases ...............................................................21 Operation DASGIP Control ..............................................76 Pumps .............................................................55 Power cord ........................................................... 35 Ordering information .........................................113 Pressure reducer .................................................. 40 Original packaging .............................................102 Process View ........................................................ 80 Other applicable documents ..................................5 Product liability .................................................... 18 Overhead drives connect ............................................................38 Connecting sockets .........................................30 Profile Constant Value ............................................... 84 Data Table ....................................................... 84 Function ......................................................... 84 Overhead Stirrer Assembly .........................................................72 Power supply ....................................................... 29 External .......................................................... 35 Internal ............................................... 30, 33, 35 Technical data .............................................. 105 Profile Source ...................................................... 83 Overlay .................................................................40 Program Start ...................................................... 75 Overpressure ........................................................32 Pt connector ....................................................... 112 P Pt100 temperature sensor ................................... 39 Packaging ...........................................................102 Pump head tubing ......................................... 49, 50 Calibrate ......................................................... 54 Deformation .................................................... 53 Insert .............................................................. 52 Parameter Definitions .......................................................82 Sensor .............................................................82 Vessel ..............................................................81 Particle filter .........................................................40 pump head tubing replace ............................................................ 99 125 Pump tubing Cleaning ..........................................................95 Pumps Continuous feeding .........................................56 Manual operation ............................................55 Short feeding 56 .................................................. Speed limits ....................................................51 Typical working ranges ...................................51 R Redox cable ..........................................................39 Redox connector ................................................112 Redox Sensors ......................................................64 Connecting sockets .........................................33 Handling instructions ......................................59 Replacement of electrolyte ...................................97 Return delivery information ...............................104 Rx/Tx ....................................................................29 Stas 200 plug 3 ....................................................... 5 State ..................................................................... 29 Sterilize Feed tubing .................................................... 54 Sensors ........................................................... 57 Storage ......................................................... 65, 102 Submerged .............................................. 15, 31, 40 Surfaces Cleaning ......................................................... 95 Symbol Colors ............................................................. 79 Controller ........................................................ 83 Vessel ............................................................. 81 Symbols Danger .............................................................. 6 Explanation ....................................................... 7 Software ......................................................... 79 S System components Delivery package ............................................ 14 Safety instructions ................................................17 T Save User Chart Settings ......................................87 Sensor membrane ..........................................61, 62 Sensors Calibrate ..........................................................58 Connect ...........................................................39 Handling instructions ......................................60 Sterilize ...........................................................57 Tab Data ................................................................ 84 Graph .............................................................. 84 Navigation ...................................................... 88 Type ................................................................ 84 Technical data .................................................... 105 Power supply ................................................ 105 Serial interfaces ....................................................28 Telescopic rails .................................................... 68 Serial number .......................................................33 Temperature control ............................................ 43 Service ............................................................96, 97 Temperature control actuator .............................. 44 Set-point Profile Definition ........................................................83 Temperature sensor ....................................... 23, 70 Connector ..................................................... 112 Port - MP8-pHpO ............................................ 33 Settings Alarm ...............................................................84 Shipment ..............................................................96 Software Process View ...................................................80 Summary View ................................................78 Symbols ...........................................................79 Splitter Function .............................................15, 42 Temperature sensors Connect .......................................................... 38 Connecting sockets - Base Unit ..................... 30 Port MP8-pHpO .............................................. 33 Terminator connector .......................................... 36 Top-heaviness Bioreactors ..................................................... 47 126 Transport ............................................................102 Typical workflow ..................................................73 U USB cable .............................................................36 USB port ...............................................................36 User profile ...........................................................17 V Ventilation grating ................................................30 Ventilation slots ....................................................33 Vessel Parameters ......................................................81 W Warning symbols 6 Devices ......................................................24, 25 Warnings Gas supply .......................................................32 Weight ................................................................106 Workflow ..............................................................73 New .................................................................77 Working Area .......................................................76 Workplace Requirements ..................................................27 Evaluate your manual Give us your feedback. www.eppendorf.com/manualfeedback Your local distributor: www.eppendorf.com/contact Eppendorf AG · 22331 Hamburg · Germany · eppendorf@eppendorf.com www.eppendorf.com
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