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In touch with life®

We Know Bioprocessing

Parallel Cultivation of Microorganisms usingSingle-use Mini BioreactorsBio Base NWE Workshop & CLIB-Forum Scale-up & Scale-down

Creative Campus Monheim, Germany | April 3, 2014 | Sebastian Kleebank

> Introduction

> Scalable Systems

> Single-use options

> Software solutions

> Summary> Summary

4

Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors

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

Introduction

> Platform technology for industry leaders in North America, Europe and Asia

> An company since Jan 1st, 2012

5Eppendorf AG | Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors | Sebastian Kleebank | April 3, 2014

Introduction

DASGIP® – Part of Eppendorf Group

Manual pipettes Electronic pipettes Pipette tips Pipette calibration Bottle-top dispensers

CombitipsDispensersAutomated pipetting

Han

dli

ng

Liq

uid

Han

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ng


Single-use vessels CO2 incubators Biological shakers Microinjectors Microinjectors Microcapillaries ElectroporatorsFermentors/ bioreactors

CentrifugesUltra-low temp-erature freezer

Mixers Spectrometers,Photometer and cuvettes

Vacuum concentrators PCR devices PCR and deepwell plates

Tubes

Sam

ple

Han

dli

ng

Cell

Han

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ng

History of Bioprocessing at Eppendorf

Introduction

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


1946 2007 2012 20131991

Foundation of DASGIP GmbH

1945

Foundation of New

Brunswick™ Scientific Co

Eppendorf AG Bioprocess

Product Lines:

New Brunswick

DASGIP

DASware®

BioBLU®

> 2700 employees in more than 25 countries

> North American, European, and Asian Headquarters

Introduction

Eppendorf – There When You Need Us

> Global sales and service

> Training and application labs throughout the world


What is located in Juelich?

Introduction

> Eppendorf Bioprocess Center:

PAN European Sales and Service Team for

New Brunswick & DASGIP bioprocess product

lines

> DASGIP GmbH:


> DASGIP GmbH:

Competence Center for Small and Parallel

Bioreactors/Fermentors

Competence Center for Advanced Interfacing

Our 3S Value Proposition

Introduction

Scalable Systems

Comprehensive

Eppendorf bioprocess

systems cover a wide range

of working volumes in

Single-Use Options

Ready-to-Use

Ready-to-use single-use

bioreactors of different

volumes complement the

Software solutions

Innovative

Comprehensive software

packages offer innovative

solutions for advanced


of working volumes in

autoclavable, single-use and

SIP.

scale range. process control and

information management.

> Introduction

> Scalable systems



> Summary> Summary

11


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)

Scalable systems


R&D and Scale DownProduct/Process

DevelopmentProduct/Process

DevelopmentPilot Scale Production

DASGIP® Product Lines

> DASbox Mini Bioreactor System

> DASGIP Parallel Bioreactor

Systems

Scalable systems

Systems

> DASGIP Control and DASware

Solutions





DASbox® Mini Bioreactor System

Optimal for screening, characterization and optimization

Scalable systems





Mini Bioreactor Solutions

DASbox® Parallel Mini Bioreactor System

> Operation of 4, 8, 12 or more vessels in parallel

> Working volumes of 60 - 250 mL

> Designed for cell culture and microbial applications

Scalable systems

applications

> Glass and single-use vessel options

> Optimal tool for DoE and scale-down approaches





DASbox® – Features

Scalable systems

Multifunctional storage unit

Local Display

Integrated Gas mixing

Feeding & Monitoring module

16

module

Liquid free temperaturecontrol unitDetachable media

bottle container and bioreactor

carrier

Eppendorf AG | Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors | Sebastian Kleebank | April 3, 2014

DASbox® Mini Bioreactor

> Glass body & stainless steel head plate

w/easy to handle screw cap

> Working volume 60 – 250 mL (H/D 2:1),

Total volume 350 mL

> Overhead drive using

Scalable systems

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)


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

Scalable systems

cultivation, cell culture, and stem cell applications

> Glass and single-use vessel options

> Highly flexible due to modular design of monitoring and control units





Benefits

> Small footprint

> Modular design

> Effective operation

Scalable systems

> Full integration

> Easy to handle





Basic Technical Features

> Individual temperature and

agitation control

> Up to 6 liquid feeds per vessel

> pH control with CO2, acid and/or

base addition

Scalable systems

> DO control by stirring speed, oxygen

concentration and/or gas flow rate

> Redox/ORP monitoring and control

> Gassing to sparge and/or overlay





Advanced Technical Features

> Miniature peristaltic pumps using variable

speed drives

> TMFC or rotameter/solenoid gassing

> O2 and CO2 exhaust analysis with online OTR,

CTR and RQ calculation

Scalable systems

> Overhead drive or magnetic stirring

> Level control or anti foam sensor

> Optical density measurement

> Illumination using LED light sticks





Bioreactor Designs

> Mammalian cell culture

> suspension

> adherent

> Microbial fermentation

> aerobic

Scalable systems

> aerobic

> anaerobic

> Phototropic cells and organisms

> Stem cells





Scale-DownExample: E. coli Fermentation

Scalable systems

Scale-Down of a 2 L amino acidfermentation process

23

Cell dry weight (X), product concentration (P) and specific product yield (YP/X), each normalized to the 2 L system

Eppendorf Application Note No. 290


Scalability – BiomassParallel E. coli K12 cultivationDry cell weight [g/L] , 22.4 h ± 1.0 h after inoculation

Scalable systems

45,0

50,0


33,930,9 32,4 31,8

0,0

5,0

10,0

15,0

20,0

25,0

30,0

35,0

40,0

SR0250ODLSn = 3

SR1000ODLSn = 4

BioBLU 0.3fn = 4

BioBLU 1fn = 4

dry

cell w

eig

ht

[g/L]

> Introduction

> Scalable systems



> Summary> Summary

25


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 3rd

Single-use options

Bioreactor Systems and CelliGen® BLU or 3rd

party systems

> Non-invasive or industry standard sensor technology





BioBLU® – A Growing Family

> Proven performance and scalability of stirred-tank design

Single-use options

Combining polymer expertise and bioprocess know-how Eppendorf offers the industry’s most complete portfolio of single-use bioreactors for RD, PD & pilot scale!

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 3rd party bioprocess systems


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

Single-use options

> Cell culture, microbial and packed-bed versions

available

> Working volume:

> 65 - 250 mL (BioBLU 0.3f)

> 100 - 250 mL (BioBLU 0.3c)

> 250 mL (BioBLU 0.3p)


Full featured

Liquid-free exhaust condenser (Peltier)

BioBLU® 0.3 – Features

Single-use options

Gas inlet to submersed and headspace gassing

Magneticallycoupled

drive assembly

Rigid wallsingle-use stirred-

tank bioreactor

Full featuredhighly integrated head plate

Heating and cooling by liquid-free DASbox temperature control


BioBLU® 1 Single-Use Vessels

> Bench scale single-use vessel


> Designed for DASGIP Bioblock providing industry

standard control capabilities

> Cell culture and microbial versions available

Single-use options

> Cell culture and microbial versions available

> Working volume:

> 250 mL – 1.25 L (BioBLU 1f)

> 320 mL – 1.25 L (BioBLU 1c)


BioBLU® 5 Single-Use Vessel

> Bench scale single-use vessel


> Parallel operation and calibration with the

DASGIP Parallel Bioreactor System

> Pitched blade (BioBLU 5c) and packed-bed

Single-use options

> Pitched blade (BioBLU 5c) and packed-bed

(BioBLU 5p) versions available

> Working volume: 1.25 – 3.75 L


Scalability – BiomassParallel E. coli K12 cultivationDry cell weight [g/L] , 22.4 h ± 1.0 h after inoculation

Single-use options

45,0

50,0


33,930,9 32,4 31,8

0,0

5,0

10,0

15,0

20,0

25,0

30,0

35,0

40,0

SR0250ODLSn = 3

SR1000ODLSn = 4

BioBLU 0.3fn = 4

BioBLU 1fn = 4

dry

cell w

eig

ht

[g/L]

> Introduction

> Scalable systems



> Summary> Summary

33


More than SCADA

Eppendorf offers a flexible range of bioprocess software solutions for Supervisory Control and Data Acquisition (SCADA) and Data Management

Software solutions


DASGIP ControlSophisticated Monitoring and Control Capabilities

BioCommandConfigurable Control Strategies,

Data logging , Alarms etc.

DASwareComprehensive Data and Information Management

New Brunswick RPCAdvanced Local Control

Comprehensive Bioprocess Information Management

DASware®

Software solutions

Remote monitoring and control via PC, Notebook, Netbook or with the DASGIP iApp via iPhone®, iPod® and iPad®

Applies Design of Experiment (DoE)



DevelopmentProcess


Seamless integration of external lab devices for process automation and feedback control loops

Integration into process control systems and corporate historians

Comprehensive and user-friendly information management solution for bioprocessing

Integration of different bioreactor control units - the gateway to benefit from DASware functionality on 3rd

party bioreactor controllers

DASGIP® Control –At the Core of All DASGIP® Systems

Software solutions

In situ sensors

Point of Measurement TrendingPoint of control

Supervisory Control and Data Acquisition (SCADA)

Control strategies & automation

Historian

Bioprocessanalytical

instruments

Recipe management


More than SCADA

> Recipe management:

load & store recipes and

configurations at different sites

> Integrated historian with 70+ data

tracks per bioreactor

Software solutions

tracks per bioreactor

> Integrated alarms & events

> Highly configurable control strategies

with data driven control decisions


Fully Integrated Workflow

Software solutions

Formulate recipe at first system/facility

Send recipe to second system/facility (via email or DASware discover)

Point

38

system/facility (via email or DASware discover)

Load and execute recipeCollect online and offline data

Send data to first system/facility for analysis(via email or DASware discover)

Click

Grow


> Introduction

> Scalable systems



> Summary> Summary

39


Culture

PassageSeed

Fermentors and Bioreactors

Freezers

Manual Liquid Handling

Centrifugation

Shakers and CO2 incubators

Summary

The Right Tools, Every Step of the Way

Manipulate

Analyze

Store

Automated Liquid Handling

Freezers

Analysis

Services

Transfection

Eppendorf AG | Parallel Cultivation of Microorganisms using Single-use Mini Bioreactors | Sebastian Kleebank | April 3, 2014 40

Scalable Systems –Total Upstream Solutions

Summary

R&D and Scale Down

Process Development


Pilot/ Production

Process Development

We Know BioprocessingThank you

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 Eppendorf logo, BioBLU®, and In touch with life® are registered trademarks of Eppendorf AG, Hamburg, Germany. New Brunswick™ is a trademark of Eppendorf AG, Hamburg, Germany.

All rights reserved, including graphics and photos. Copyright © 2014 by Eppendorf AG.


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