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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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