Automating the Bioprocessing Workflow

Transcription

1 24 September 2014 Automating the Bioprocessing Workflow Daniel Leach Senior Application Scientist

2 Biotherapeutic Workflow

3 Design of Experiment Generate Experimental Conditions & Experimental Design Execute Experiment Determine Experimental Results Analyze Experimental Data

4 Design of Experiment Generate Experimental Conditions & Experimental Design Execute Experiment Determine Experimental Results Analyze Experimental Data

5 Design of Experiment Binding Capacity Study Experimental design Doehlert design Factors ph: 4, 4.5, 5, 5.5, 6 NaCl: 0, 50, 100 mm Response Fab (Biacore data) A B C D E F G H ph NaCl (mm) ph NaCl Doehlert design ph Replicates Capacity = q = V liq /V resin [ (c o -c FT1 ) + (c o -c FT2 ) + (c o -c FT3 ) ] Data Supplied by GE Healthcare

6 Design of Experiment Binding Capacity Conditions 500mM 464mM 427mM 390mM 355mM 318mM 282 mm 245 mm 209 mm 172 mm 136 mm 100 mm

7 Design Experimental Conditions

8 Design of Experiment Generate Experimental Conditions & Experimental Design Execute Experiment Determine Experimental Results Analyze Experimental Data

9 Record Phase Names & Buffer Type used

10 Generate Phase Conditions Create Extensive Report

11 Generate Load Conditions vary flow per tip/column

12 Generate Experimental Conditions

13 Design of Experiment Generate Experimental Conditions & Experimental Design Execute Experiment Determine Experimental Results Analyze Experimental Data

14 Tecan Modules for Phynexus Robotic Manipulator Arm (RoMa) Liquid Handling Arm (LiHa) Plate and rack transport 8 channel Transport to devices Disposable tips Multi Channel Arm (MCA) 96-channel pipetting head Disposable tips Required for Gel Filtration rack Either Or

15 Design of Experiment Generate Experimental Conditions & Experimental Design Execute Experiment Determine Experimental Results Analyze Experimental Data

16 Experimental Example : Method Hydridoma Supernatant Purification Fully automated purification 96 supernatants processed in less than 15 minutes Phases : Equilibration, Capture, Wash 1, Wash 2 and Enrichment Equilibration : 1 cycle 8.3 ul/s flow rate Capture : 8 cycles 4.0 ul/s flow rate Wash 1 & 2 : 1 cycle 8.3 ul/s flow rate Enrich : 8 cycles 4.0 ul/s flow rate Data from Tecan Application Note True parallel protein purification with PhyTip columns using the Freedom EVO and MultiChannel Arm 96

17 Experimental Analytics Protein Quantification Fully automated quantification 280nm read of every fraction collected Determine total volume collected for each purification

18 Experimental Example : Results Hydridoma Supernatant Purification 24 identical samples purified with PhyTips Simultaneous processing of PhyTips with MCA96 pipetting arm Protein Recovery : Over 74% % CV Eluted Volume : 5% % CV Concentration : 10% Conclusions PhyTip columns perform best using back and forth aspirate/dispense cycles for optimal capture, wash and elution PhyTips used in conjunction with the Freedom EVO are capable of achieving reproducible purification results The Freedom EVO is suitable for processing PhyTip columns, enabling complete automation of sample preparation and protein purification Data from Tecan Application Note True parallel protein purification with PhyTip columns using the Freedom EVO and MultiChannel Arm 96

19 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

20 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)

21 Experimental Analytics Plate based ELISA Kit based ELISA s for specific antibody/proteins Onboard sample dilutions Complete walk-away High throughput analysis

22 Experimental Analytics Biomolecular Interactions ForteBIO integration Dip & Read Complete walk-away High throughput analysis

23 Experimental Analytics Protein Analysis Electrophoretic analysis Lab chip integration Fully integrated sample preparation and measurement with GXII workflow

24 Experimental Analytics Chromatographic Analysis Prepare fractions for further chromatographic analysis

25 Design of Experiment Generate Experimental Conditions & Experimental Design Execute Experiment Determine Experimental Results Analyze Experimental Data

26 Data Analysis ASSIST (interpolated values) NaCl (mm) ph Data Supplied by GE Healthcare

27 Design the Next Experiment

28 Thank you for your attention Tecan Group AG is making all efforts to include accurate and up-to-date information into this presentation. Yet, it cannot be ruled out that omissions or errors might have occurred. Therefore, Tecan Group AG cannot make any representations or warranties, expressed or implied, as to the accuracy or completeness of the information provided in this presentation. Changes in this presentation can be made 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. Tecan Trading AG, Switzerland, all rights reserved.