PreDictor platform and Tecan automation —effi cient

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

4

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

5

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.

6

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.

7

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.

8

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.

9

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

10

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.

11

Freedom EVO —specifications

Ordering information

PreDictor plates are available with Capto, MabSelect, and

Q and SP Sepharose™ Fast Flow chromatography media

For details visit:


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:



For local office contact information, visit

www.tecan.com/contactTecan Switzerland Seestrasse 103 8408 Männedorf ZH Switzerland


For local office contact information, visit

www.gelifesciences.com/contactGE Healthcare Bio-Sciences AB Björkgatan 30 751 84 Uppsala Sweden


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

Documents

PreDictor platform and Tecan automation —effi cient