predictor platform and tecan automation —effi cient
TRANSCRIPT
GE Healthcare
PreDictor platform and Tecan automation—effi cient solutions for process development
In the development of biopharmaceuticals, time-to-clinic
and time-to-market are key factors for the success of any
project. That’s why efficient process development is a
crucial component of the overall project plan. Screening
for optimal conditions can be time-consuming and tedious
parts of the process.
GE Healthcare and Tecan are working together to ease
your task in drug development by providing the tools
you need to succeed. With over 50 years of experience,
we have recognized expertise in our respective fields
of process development and laboratory automation.
We are collaborating to provide robust solutions for
high-throughput process development so that you can
concentrate on drug development.
More information • Less time • Full automation
Welcome to a new era of process development.
GE Healthcare and Tecan— working together to speed up process development
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Automation
The development of protocols for parallel screening of
chromatographic conditions is essential to decrease the
time required for process development. Protocols reduce
overall time-to-clinic and allow for more drug candidates
to be screened in a shorter time.
The development of automated workflows increases
throughput and enables methods to be performed with a
high degree of robustness. Automated work-fl ows enable
hands-off operation and eliminate human error, resulting
in uncompromising experimental reproducibility and
consistency among replicates.
This brochure presents proven automated solutions
for high-throughput process development. We present
some applications and the excellent reproducibility that
underlies all the featured products.
For more information, please visit:
www.gelifesciences.com/predictor
www.tecan.com/predictor
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PreDictor
PreDictor™ 96-well plates support high-throughput
process development (HTPD) by allowing parallel
screening of chromatographic conditions. They can
be operated with both centrifugation and vacuum
protocols, manually or in automated workfl ows.
PreDictor plates are prefilled with GE Healthcare
BioProcess™ chromatography media and have been
developed to evaluate conditions for binding, wash
or elution, or to determine adsorption isotherms.
Experiments are performed based on batch separation,
which eliminates errors induced by non-optimal formats
or packing procedures. Data generated with PreDictor
plates show good correlation with data generated
in chromatography columns, making the plates an
excellent tool for the initial screening of process
conditions.
Assist software for PreDictor plates
To facilitate the high-throughput method, we have
developed Assist software to support PreDictor plates.
The software guides you through experimental setup
and workfl ow to data management and subsequent
evaluation.
PreDictor plates and the HTPD workfl ow result in:• Shorter time-to-clinic
• More process information in less time
• Reduced sample consumption
PreDictor plates are available with our latest chromato-
graphy media families Capto™ and MabSelect™. The
plates are also available with combinations of media for
screening purposes.
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Fig 1. Schematic diagram of an experimental setup using different incubation times on a PreDictor plate. The sample to be incubated the longest time is added first, while the sample to be incubated the shortest time is added last. At the end of the shortest incubation time the phases are separated simultaneously in all wells using an appropriate technique, e.g., vacuum filtration or centrifugation.
Sampleaddition
Sampleaddition
Sampleaddition
Sampleaddition
Sampleaddition
StartReal time
End
StartEndIncubation time
Sim
ulta
neou
s se
para
tion
of p
hase
s an
dsu
bseq
uent
con
cent
ratio
n m
easu
rem
ents
Flow
thro
ugh
Elut
ion
PreDictor data used for the prediction of column behavior
Determination of dynamic binding capacity (DBC) is
very important during process development because it
describes the binding capacity under different operating
conditions. Traditionally, DBC determination has involved
tedious and time consuming experiments performed via
column chromatography.
In the following example, the DBC of a monoclonal
antibody (MAb) to MabSelect SuRe™ was predicted from
data generated on PreDictor MabSelect SuRe, 6 µl.
Prediction of dynamic binding capacity
The workflow was fully automated on a Tecan Freedom
EVO® 200 robotic workstation. The experimental setup
included four different sample concentrations (Co, Co/2,
Co/4, Co/8) and six different incubation times. Figure 1
shows the basic principle for time-dependent studies in
96-well plates.
Concentrations of MAb in the eluate pool were
determined spectrophotometrically. Static binding
capacities were calculated and plotted as a function
of incubation time. Four uptake curves were obtained
(Fig 2), one for each initial protein concentration tested.
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30 60 90 120 Time (min)
A
B
Preparation time
Hands-on time during process
Fig 2. Uptake curves recorded at different initial concentrations are depicted as temporal changes in binding capacity on MabSelect SuRe. Error bars represent standard deviations calculated from four replicates.
0
5
10
15
20
25
30
35
40
45
50
0 10 20 30 40 50 60 70
Co
Co/2
Co/4
Co/8
Incubation time (min)
Cap
acity
(g/l
)
0
0.2
0.4
0.6
0.8
1
1.2
0 2 4 6 8 10 12Residence time (min)
Nor
mal
ized
DBC
Plate
Column
Fig 3. A comparison of predicted DBC at various residence times (0.5, 1, 2, 4, 8, and 10 min) based on results from PreDictor plates with DBC obtained by frontal analysis in column experiments (1 ml HiTrap™ columns).
Conclusion
The PreDictor 96-well plate is a time-saving and
accurate tool for the determination of binding
capacity. Data can be used for the prediction of
dynamic binding capacity in columns.
Automation on Freedom EVO enables hands-off
operation and ensures consistent results.
The data was used to predict the DBC shown in
Figure 3. There was a good correlation between the
predicted DBC and the DBC on a column.
Fig 4. Time required to generate the time-dependent protein uptake curves described above by automated (A) and manual (B) process. Preparation time is shown in blue and subsequent hands-on time in red.
The time required to generate the time-dependent
protein uptake curves in PreDictor plates by automated
and manual processes are shown in Figure 4.
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Consistent well-to-well and plate-to-plate performance
depends on both the reproducibilty of the PreDictor
plates and the liquid handling capability of the robot.
Figure 5 shows the binding capacities determined in
a 2 µl PreDictor Capto S plate after overloading it with
protein. The relative standard deviation (RSD) calculated
over several plates was 4.5%. This low variation in
binding capacity clearly demonstrates the low variation
in medium volume between wells when compared to
the variability of the UV assay (RSD = 3%).
Figure 6 highlights the reproducibility of the processing
of PreDictor plates on a Tecan Freedom EVO robotic
workstation equipped with a Te-Shake™ and a
Te-VacS™ vacuum separation module. A PreDictor
Highly reproducible results
0
5
10
15
20
25
30
2 µl 6 µl 20 µl 50 µl
Medium volume in wells (µl)
Nor
mal
ized
med
ium
pre
senc
e
First occasion Second occasion Third occasion
Fig 6. Excellent reproducibility in pipetting and liquid handling on Tecan Freedom EVO. The amount of media pore volume marker was recalculated as pore liquid. Data from each run is shown as one bar and error bars correspond to 95% confidence intervals. The three consecutive runs for each media volume are shown side by side.
Fig 5. Reproducibility of capacities for chymotrypsin (60 min incubation) determined in a 2 µl PreDictor Capto S plate. The solid red line corresponds to the average value, dashed lines denote ± 2 standard deviations. Columns 1 to 12 on the PreDictor plate are denoted by the alternating colored bars, indicating the absence of any significant edge effects.
0
50
100
150
200
* PreDictor plates are delivered with these different volumes.
plate prefilled with 16 wells each of 2, 6, 20, and 50 µl
chromatography media*, was saturated with a media
pore volume marker. The pore volume marker was then
washed away with 3 × 100 µl water (using vacuum for
liquid removal), collected, and measured. The procedure
was repeated twice, resulting in three consecutive runs
on the same plate.
Altogether, these results demonstrate the excellent
reproducibility inherent in both PreDictor plate
filling volumes and also within PreDictor plates on
Freedom EVO.
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Freedom EVOware software—simple, intuitive, fl exible
Freedom EVOware softwareFreedom EVOware® is an easy-to-use software for
controlling robotic workstations. It allows complete
automation of the entire process, starting with the
preparation of buffers from stock solutions to result
analysis in a Tecan reader.
Freedom EVOware offers:• An intuitive graphical user interface with a drag-and-
drop feature for every development step
• Precise control of incubation times for increased repeatability
• Sample tracking and documentation
• Wizards for easy creation of advanced workfl ows
• Comprehensive and precise control of Tecan devices such as Te-VacS and Te-Shake
• Integration of a variety of third party devices, such as centrifuges
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Accessories dedicated to PreDictor
We provide dedicated accessories for the robust and
reliable processing of PreDictor plates on Freedom EVO.
These include a Holder fi lter paper and a Plate shaker
PreDictor frame for reducing the risk of leakage from
individual wells.
Throughout the protocol, droplets beneath the fi lter
have to be removed from the bottom of the plate.
PreDictor plates are therefore blotted onto a blotting
paper held in place by the Holder filter paper. The
The robotic manipulator arm with centric grippers is placing a PreDictor plate onto a microtiter plate on the Plate shaker PreDictor frame.
The robotic manipulator arm with centric grippers blots a PreDictor plate on a blotting paper held in place by the Holder fi lter paper.
holders can be handled by the robotic manipulator
arm and they can be stored in hotels, carousels, and
on microtiter plate carriers on the worktable.
The Plate shaker PreDictor frame is used during
incubation on Te-Shake. It holds up to two PreDictor
plates placed on top of a conventional microtiter
plate that is used to avoid direct contact between
the bottom of the PreDictor plate and the surface
underneath it.
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Freedom EVO —specifications
Ordering information
PreDictor plates are available with Capto, MabSelect, and
Q and SP Sepharose™ Fast Flow chromatography media
For details visit:
www.gelifesciences.com/predictor
General hardware featuresRobotic arms Liquid handling arms, robotic manipulators, several multichannel options
Tip configuration 1, 2, 4, 8, tips, various conbinations of application-oriented tip types
Tip types Standard (Teflon™ coated stainless steel) and disposable tips with or without
filter (10/200/1000 µl); low-volume tips for high-density format applications
Syringe sizes 25/50/250/500/1000/2500/5000 µl
Liquid handling featuresVolume range 0.5 µl–5000 µl
Pipetting precision Volume Standard tips Disposable tips
200 µl 1000 µl
10 µl ≤ 3.0% ≤ 3.0% —
100 µl ≤ 0.5% ≤ 0.5% ≤ 1.0%
For more information, please visit:
www.gelifesciences.com/predictor
www.tecan.com/predictor
For local office contact information, visit
www.tecan.com/contactTecan Switzerland Seestrasse 103 8408 Männedorf ZH Switzerland
www.tecan.com/predictor
For local office contact information, visit
www.gelifesciences.com/contactGE Healthcare Bio-Sciences AB Björkgatan 30 751 84 Uppsala Sweden
www.gelifesciences.com/predictor
GE, imagination at work, and GE monogram are trademarks of General Electric Company.
BioProcess, Capto, HiTrap, MabSelect, MabSelect SuRe, PreDictor, and Sepharose are trademarks of GE Healthcare companies.
All third party trademarks are the property of their respective owners.
© 2009 General Electric Company—All rights reserved. First published Feb. 2009
All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information.
GE Healthcare Bio-Sciences AB Björkgatan 30, 751 84 Uppsala Sweden
GE Healthcare Europe, GmbH Munzinger Strasse 5, D-7911 Freiburg Germany
GE Healthcare Bio-Sciences Corp. 800 Centennial Avenue, P.O. Box 1327 Piscataway, NJ 08855-1327 USA
GE Healthcare Bio-Sciences KK Sanken Bldg., 3-25-1, Hyakunincho Shinjuku-ku, Tokyo 169-0073 Japan
Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this publication, however, it is possible that omissions or errors might have occurred. Tecan Group Ltd. cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy or completeness of the information provided in this publication. Changes in this publication can be made at any time without notice. All mentioned trademarks are protected by law.
For technical details and detailed procedures of the specifications provided in this document please contact your Tecan representative. This brochure may contain reference to applications and products which are not available in all markets. Please check with your local sales representative.
The applications described here are for research use only. Not for use in human clinical or diagnostic procedures. Freedom EVO and Freedom EVOware are registered trademarks, and Te-Shake and Te-VacS are trademarks of Tecan Group Ltd., Männedorf, Switzerland.
© 2009, Tecan Trading AG, Switzerland, all rights reserved
28-9403-60 AA 03/2009