We Know Bioprocessing
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1 In touch with life
2 We Know Bioprocessing
3 Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors Bio Base NWE Workshop & CLIB-Forum Scale-up & Scale-down Creative Campus Monheim, Germany April 3, 2014 Sebastian Kleebank
4 > Introduction > Scalable Systems > Single-use options > Software solutions > Summary Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors 4
5 Introduction DASGIP Information and Process Technology GmbH Excellence in hardware & software solutions > Founded in 1991 as a computer and software engineering company > Developing benchtop bioprocessing solutions since 1994 > Commitment to customers and product for more than 20 years > Platform technology for industry leaders in North America, Europe and Asia > An company since Jan 1st,
6 Introduction DASGIP Part of Eppendorf Group Sample Handling Cell Hand dling Liquid Handling Manual pipettes Electronic pipettes Pipette tips Pipette calibration Automated pipetting Dispensers Combitips Bottle-top dispensers Fermentors/ bioreactors Centrifuges Single-use vessels CO 2 incubators Biological shakers Microinjectors Microinjectors Microcapillaries Electroporators Mixers Vacuum concentrators PCR devices Ultra-low temperature freezer Spectrometers, Photometer and cuvettes PCR and deepwell plates Tubes 6
7 Introduction History of Bioprocessing at Eppendorf 70 years of experience in the life science market Foundation of Eppendorf- Netheler-Hinz GmbH Acquisition of New Brunswick Scientific Co Acquisition of DASGIP GmbH Foundation of New Brunswick Scientific Co Foundation of DASGIP GmbH EppendorfAG Bioprocess Product Lines: New Brunswick DASGIP DASware BioBLU 7
8 Introduction Eppendorf There When You Need Us > 2700 employees in more than 25 countries > North American, European, and Asian Headquarters > Global sales and service > Training and application labs throughout the world 8
9 Introduction What is located in Juelich? > Eppendorf Bioprocess Center: PAN European Sales and Service Team for New Brunswick & DASGIP bioprocess product lines > DASGIP GmbH: Competence Center for Small and Parallel Bioreactors/Fermentors Competence Center for Advanced Interfacing 9
10 Introduction Our 3S Value Proposition Scalable Systems Comprehensive Eppendorf bioprocess systems cover a wide range of working volumes in autoclavable, single-use and SIP. Single-Use Options Ready-to-Use Ready-to-use single-use bioreactors of different volumes complement the scale range. Software solutions Innovative Comprehensive software packages offer innovative solutions for advanced process control and information management. 10
11 > Introduction > Scalable systems > Single-use options > Software solutions > Summary Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors 11
12 Scalable systems Bioreactor Solutions > Glass and stainless steel bioreactors with working volumes of 60 ml 2400 L > Single-use bioreactors with working volumes of 65 ml 1.25 L (microbial application) and 100 ml 40 L (cell culture) R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 12
13 Scalable systems DASGIP Product Lines > DASbox Mini Bioreactor System > DASGIP Parallel Bioreactor Systems > DASGIP Control and DASware Solutions R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 13
14 Scalable systems DASbox Mini Bioreactor System Optimal for screening, characterization and optimization R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 14
15 Scalable systems Mini Bioreactor Solutions DASbox Parallel Mini Bioreactor System > Operation of 4, 8, 12 or more vessels in parallel > Working volumes of ml > Designed for cell culture and microbial applications > Glass and single-use vessel options > Optimal tool for DoE and scale-down approaches R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 15
16 Scalable systems DASbox Features Multifunctional storage unit Integrated Gas mixing Local Display Feeding & Monitoring module Detachable media bottle container and bioreactor carrier Liquid free temperature control unit 16
17 Scalable systems DASbox Mini Bioreactor > Glass body & stainless steel head plate w/easy to handle screw cap > Working volume ml (H/D 2:1), Total volume 350 ml > Overhead drive using miniature lip seal assembly with Rushton and/or marine impellers > Liquid free exhaust condenser > Utilizing off-the-shelf ph and DO sensors (120mm long, 12mm diameter) 17
18 Scalable systems Small-and Bench scale Bioreactor Solutions DASGIP Parallel Bioreactor Systems > Parallel operation of 4, 8 or more vessels > Working volumes of 35 ml 3.8 L > Designed for fermentation, phototrophic cultivation, cell culture, and stem cell applications > Glass and single-use vessel options > Highly flexible due to modular design of monitoring and control units R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 18
19 Scalable systems Benefits > Small footprint > Modular design > Effective operation > Full integration > Easy to handle R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 19
20 Scalable systems Basic Technical Features > Individual temperature and agitation control > Up to 6 liquid feeds per vessel > ph control with CO2, acid and/or base addition > DO control by stirring speed, oxygen concentration and/or gas flow rate > Redox/ORP monitoring and control > Gassing to sparge and/or overlay R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 20
21 Scalable systems Advanced Technical Features > Miniature peristaltic pumps using variable speed drives > TMFC or rotameter/solenoid gassing > O 2 and CO 2 exhaust analysis with online OTR, CTR and RQ calculation > Overhead drive or magnetic stirring > Level control or anti foam sensor > Optical density measurement > Illumination using LED light sticks R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 21
22 Scalable systems Bioreactor Designs > Mammalian cell culture > suspension > adherent > Microbial fermentation > aerobic > anaerobic > Phototropic cells and organisms > Stem cells R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 22
23 Scalable systems Scale-Down Example: E. coli Fermentation Scale-Down ofa 2 L aminoacid fermentation process Eppendorf Application Note No. 290 Cell dry weight (X), product concentration (P) and specific product yield (Y P/X ), each normalized to the 2 L system 23
24 Scalable systems Scalability Biomass Parallel E. coli K12 cultivation Dry cellweight [g/l], 22.4 h ± 1.0 h after inoculation 50,0 45,0 dry cell weight [g/l] 40,0 35,0 30,0 25,0 20,0 15,0 10,0 5,0 0,0 33,9 SR0250ODLS n = 3 30,9 32,4 31,8 SR1000ODLS n = 4 BioBLU 0.3f n = 4 BioBLU 1f n = 4 24
25 > Introduction > Scalable systems > Single-use options > Software solutions > Summary Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors 25
26 Single-use options BioBLU Rigid Wall, Stirred-Tank Single- Use Vessels > Various types for cell culture and microbial applications, including packed-bed version > Working volumes of 100 ml 40 L (cell culture) and 65 ml 1.25 L (microbial applications) > To be operated with DASbox, DASGIP Parallel Bioreactor Systems and CelliGen BLU or 3 rd party systems > Non-invasive or industry standard sensor technology R&D and Scale Down Product/Process Product/Process Development Development Pilot Scale Production 26
27 Single-use options BioBLU A Growing Family Combining polymer expertise and bioprocess know-how Eppendorfoffers the industry s most complete portfolio of single-use bioreactors for RD, PD & pilot scale! > Proven performance and scalability of stirred-tank design > User-friendly set-up for rapid turnover, shorter development times and lower operating costs > USP Class VI and animal component free materials making vessels appropriate for GMP production > Adapter kits available for 3 rd party bioprocess systems 27
28 Single-use options BioBLU 0.3 Single-Use Vessels > Small scale single-use vessel > Classic stirred-tank bioreactor design > Designed for DASbox Mini Bioreactor System providing industry standard control capabilities > Cell culture, microbial and packed-bed versions available > Working volume: > ml (BioBLU 0.3f) > ml (BioBLU 0.3c) > 250 ml (BioBLU 0.3p) 28
29 Single-use options BioBLU 0.3 Features Liquid-free exhaust condenser (Peltier) Gas inlet to submersed and headspace gassing Magnetically coupled drive assembly Rigid wall single-use stirredtank bioreactor Full featured highly integrated head plate Heating and cooling by liquid-free DASbox temperature control 29
30 Single-use options BioBLU 1 Single-Use Vessels > Bench scale single-use vessel > Classic stirred-tank bioreactor design > Designed for DASGIP Bioblock providing industry standard control capabilities > Cell culture and microbial versions available > Working volume: > 250 ml 1.25 L (BioBLU 1f) > 320 ml 1.25 L (BioBLU 1c) 30
31 Single-use options BioBLU 5 Single-Use Vessel > Bench scale single-use vessel > Classic stirred-tank bioreactor design > Parallel operation and calibration with the DASGIP Parallel Bioreactor System > Pitched blade (BioBLU 5c) and packed-bed (BioBLU 5p) versions available > Working volume: L 31
32 Single-use options Scalability Biomass Parallel E. coli K12 cultivation Dry cellweight [g/l], 22.4 h ± 1.0 h after inoculation 50,0 45,0 dry cell weight [g/l] 40,0 35,0 30,0 25,0 20,0 15,0 10,0 5,0 0,0 33,9 SR0250ODLS n = 3 30,9 32,4 31,8 SR1000ODLS n = 4 BioBLU 0.3f n = 4 BioBLU 1f n = 4 32
33 > Introduction > Scalable systems > Single-use options > Software solutions > Summary Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors 33
34 Software solutions More than SCADA Eppendorfoffers a flexible range of bioprocess software solutions for Supervisory Control and Data Acquisition (SCADA) and Data Management DASGIP Control Sophisticated Monitoring and Control Capabilities DASware Comprehensive Data and Information Management New Brunswick RPC Advanced Local Control BioCommand Configurable Control Strategies, Data logging, Alarms etc. 34
35 Software solutions Comprehensive Bioprocess Information Management DASware Remote monitoring and control via PC, Notebook, Netbook or with the DASGIP iappvia iphone, ipod and ipad Seamless integration of external lab devices for process automation and feedback control loops Integration into process control systems and corporate historians Applies Design of Experiment (DoE) Comprehensive and user-friendly information management solution for bioprocessing Integration of different bioreactor control units -the gateway to benefit from DASwarefunctionality on 3 rd party bioreactor controllers R&D and Scale Down Process Product/Process Development Development Pilot Scale Production 35
36 Software solutions DASGIP Control At the Core of All DASGIP Systems Point of Measurement In situ sensors Point of control Supervisory Control and Data Acquisition (SCADA) Trending Control strategies & automation Bioprocess analytical instruments Recipe management Historian 36
37 Software solutions More than SCADA > Recipe management: load & store recipes and configurations at different sites > Integrated historian with 70+ data tracks per bioreactor > Integrated alarms & events > Highly configurable control strategies with data driven control decisions 37
38 Software solutions Fully Integrated Workflow Formulate recipe at first system/facility Point Click Grow Send recipe to second system/facility (via or DASware discover) Send data to first system/facility for analysis (via or DASware discover) Collect online and offline data Load and execute recipe 38
39 > Introduction > Scalable systems > Single-use options > Software solutions > Summary Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors 39
40 Summary The Right Tools, Every Step of the Way Shakers and CO 2 incubators Fermentorsand Bioreactors Culture Manual Liquid Handling Seed Passage Centrifugation Freezers Store Manipulate Transfection Analysis Analyze Services Automated Liquid Handling 40
41 Summary Scalable Systems Total Upstream Solutions R&D and Scale Down Process Development Pilot/ Production Process Development 41
42 We Know Bioprocessing Thank you
43 Trademarks CelliGen and BioCommand are registered trademarks of Eppendorf, Inc., Enfield, Connecticut (USA). DASGIP, DASbox and DASware are registered trademarks of DASGIP Information and Process Technology GmbH, Juelich, Germany. Eppendorf the Eppendorflogo, BioBLU, and In touch with life are registered trademarks of EppendorfAG, Hamburg, Germany. New Brunswick is a trademark of EppendorfAG, Hamburg, Germany. All rights reserved, including graphics and photos. Copyright 2014 by Eppendorf AG. 43
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