24 September 2014

Automating the Bioprocessing Workflow

Daniel LeachSenior Application Scientist

Biotherapeutic Workflow

Design of Experiment

Generate Experimental Conditions & Experimental Design

Execute Experiment

Determine Experimental Results

Analyze Experimental Data

Design of Experiment

Generate Experimental Conditions & Experimental Design

Execute Experiment

Determine Experimental Results

Analyze Experimental Data

Design of Experiment – Binding Capacity Study

pH

Replicates

A

B

C

D

E

F

G

H

121 2 3 4 5 6 7 8 9 10 11

6.0 50

5.5 100

4.0 50

4.5 0

5.5 0

4.5 100

5.0 50

5.0 50

pH NaCl (mM) NaCl100

50

0pH

4 5 6

Doehlert design

Experimental design• Doehlert design

Factors• pH: 4, 4.5, 5, 5.5, 6

• NaCl: 0, 50, 100 mM

Response• Fab (Biacore data)

Capacity = q = Vliq/Vresin × [ (co-cFT1) + (co-cFT2) + (co-cFT3) ]

Data Supplied by GE Healthcare

Design of Experiment – Binding Capacity Conditions

10

0m

M

13

6m

M

17

2m

M

20

9m

M

24

5m

M

28

2m

M

31

8m

M

35

5m

M

39

0m

M

42

7m

M

46

4m

M

50

0m

M

Design Experimental Conditions

Design of Experiment

Generate Experimental Conditions & Experimental Design

Execute Experiment

Determine Experimental Results

Analyze Experimental Data

Record Phase Names & Buffer Type used

Generate Phase Conditions – Create Extensive Report

Generate Load Conditions – vary flow per tip/column

Generate Experimental Conditions

Design of Experiment

Generate Experimental Conditions & Experimental Design

Execute Experiment

Determine Experimental Results

Analyze Experimental Data

Robotic ManipulatorArm (RoMa)

Liquid Handling Arm(LiHa)

Multi Channel Arm™(MCA)

• Plate and rack transport

• Transport to devices

• 8 channel

• Disposable tips

• 96-channel pipettinghead

• Disposable tips

• Required for GelFiltration rack

• Either • Or

Tecan Modules for Phynexus

Design of Experiment

Generate Experimental Conditions & Experimental Design

Execute Experiment

Determine Experimental Results

Analyze Experimental Data

Experimental Example : Method Hydridoma Supernatant Purification

• Fully automated purification

• 96 supernatants processed in less than 15 minutes

• Phases : Equilibration, Capture, Wash 1, Wash 2 andEnrichment

• Equilibration : 1 cycle (200uL) @ 8.3 uL/s flow rate

• Capture : 8 cycles (50uL) @ 4.0 uL/s flow rate

• Wash 1 & 2 : 1 cycle (200uL) @ 8.3 uL/s flow rate

• Enrich : 8 cycles (40uL) @ 4.0 uL/s flow rate

Data from Tecan Application Note 396301 –” True parallel protein purification with PhyTip® columns

using the Freedom EVO® and MultiChannel Arm™ 96”

Experimental Analytics

Protein Quantification

• Fully automated quantification

• 280nm read of every fraction collected

• Determine total volume collected for each purification

Experimental Example : Results

Hydridoma Supernatant Purification

• 24 identical samples purified withPhyTips

• Simultaneous processing of PhyTipswith MCA96 pipetting arm

• Protein Recovery : Over 74%

• % CV Eluted Volume : 5%

• % CV Concentration : 10%

Conclusions

• PhyTip columns perform best usingback and forth aspirate/dispense cyclesfor optimal capture, wash and elution

• PhyTips used in conjunction with theFreedom EVO are capable of achievingreproducible purification results

• The Freedom EVO is suitable forprocessing PhyTip columns, enablingcomplete automation of samplepreparation and protein purification

Data from Tecan Application Note 396301 –” True parallel protein purification with PhyTip® columns

using the Freedom EVO® and MultiChannel Arm™ 96”

Experimental Analytics

pH

• Automation manipulated pH probe

• Dip & Read approach

• Clean and Dry probe

• Automated data capture from pH probe

• Automated calibration

• Measure 96 wells in ~60 mins

Experimental Analytics

pH and Conductivity

• Measurements in different labware

• Onboard sensor cleaning and drying

• Onboard sensor calibration

• Measure 96 wells in ~30 mins (dual data)

Experimental Analytics

Plate based ELISA

• Kit based ELISA’s for specific antibody/proteins

• Onboard sample dilutions

• Complete walk-away

• High throughput analysis

Experimental Analytics

Biomolecular Interactions

• ForteBIO integration

• Dip & Read

• Complete walk-away

• High throughput analysis

Experimental Analytics

Protein Analysis

• Electrophoretic analysis

• Lab chip integration

• Fully integrated sample preparation and measurement with GXII workflow

Experimental Analytics

Chromatographic Analysis

• Prepare fractions for further chromatographic analysis

Design of Experiment

Generate Experimental Conditions & Experimental Design

Execute Experiment

Determine Experimental Results

Analyze Experimental Data

Data Analysis

4 5 60

100

50

pH

Na

Cl(

mM

)

ASSIST (interpolated values)

Data Supplied by GE Healthcare

Design the Next Experiment

Thank you for your attention

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