Environmental Impact on Liquid Handling
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1 Environmental Impact on Liquid Handling Pascal Silversmet, GSK Katie Haith, Tecan George Rodrigues, Artel 1
2 GSK : Who we are Pharmaceuticals We develop and make medicines to treat a range of conditions including respiratory diseases and HIV/AIDS 2bn packs of medicines in 2016 Vaccines We research and make vaccines for children and adults that protect against infectious diseases 833m doses of vaccines in 2016 Consumer Healthcare We make a range of consumer healthcare products in four categories: Wellness, Skin health, Oral health and Nutrition 5bn consumer healthcare products in 2016 Source: GSK 2016 Annual Report 2
3 Our global presence Saint-Amand-Les-Eaux Dresden Rixensart Moscow Ste Foy Hamilton Hamilton Wavre Marburg Nashik Tian Yuan (JV) Gödollö Shanghai Marietta Rosia/Siena Ankleshwar Singapore Manufacturing Facilities 3
4 Our global presence Saint-Amand-Les-Eaux Dresden Rixensart Moscow Ste Foy Hamilton Hamilton Rockville Wavre Marburg Nashik Tian Yuan (JV) Gödollö Shanghai Marietta Rosia/Siena Global R&D Centers Ankleshwar Singapore Manufacturing Facilities 4
5 EasyLab Global support to R&D Global automation park CAN RDC US RDC Belgium EASyLab Centre of Excellence Marburg USA RDC Italy CHN BRA users - >80 running systems - 7 countries, accross 3 continents 5
6 EasyLab : Engineering Automated Systems for Laboratory Mission Support Global R&D in : Integrate Automated Solutions into Lab Processes with a end-to-end service and harmonization mindset Project Design Phase Implementation Phase Development Validation LCM 6
7 EasyLab Expertise Main Technological Expertise Field Liquid Handling (topic of today) Sample preparation & Predilution, Serial Dilution, Plate Filling & Reagent dispensing, Aliquoting, Formulation and ph measurement, Image Analysis Cells based assay reader: Seroneutralisation, Titration, Elispot, SRD, Bactericide, Hemaglutination, (Fluorescence, multiplex, ) Automated Label printing & Sample Management Customized Robotics solution Innovative technologies : Artificial Intelligence, Augmented Reality Computer System Validation (Up to GMP level) 7
8 Liquid Handling : Precision/accuracy qualification Case study : Laval 2015 (Canada) Situation Clinical Lab equipped with 6 TECAN Liquid handling systems Systems are working days and night to produce Clinical results Ecological initiatives in 2015 to save energy Nobody in the Lab the weekend STOP HVAC (air conditioning) in lab WE Canada winter -40 C and summer +40 extreme different environment conditions Issue Suddenly, Assays performed on LH systems on Monday systematically out of range Consequence : No more testing on Monday decrease productivity & trust in automated systems After investigation : Humidity/pressure/temperature required all Monday to recover. No real human sensitivity but LH systems affected. 8
9 Liquid Handling : Environmental impact Partnership TECAN-ARTEL-GSK As global organization EASyLab transfer system/protocol to other GSK sites and countries different weather conditions and to workaround we need : Environment Automatic Liquid Handling parameters adjustment or At least monitor temperature/pressure/humidity for further investigation Partnership between ARTEL-TECAN-GSK to Modelize impact of environment on LH based on Design of Experiment approach where : GSK provided statistical design Tecan provided equipment and liquid handling expertise Artel provided environmental chamber, and conducted testing and data analysis 9
10 Tecan QC Kit xx February 2018 Katie Haith Tecan US XX
11 Freedom EVO Series Freedom EVO 150 equipped with AirLiHa, RoMa and MCA96 11
12 QC Kit components HARDWARE Calibrator plate Reader Shaker Barcode Scanner TEST MATERIALS SOFTWARE QC Kit Verification Plates QC Kit Solutions (96 well & 384 well) Freedom EVOware QC Kit Wizard 12
13 QC Kit Sample Solutions & Diluent Contain 2 dyes: Diluent and Range Solution Distinct absorbance maxima (520 & 730 nm) Different concentrations of Range (red dye) for different volume ranges Diluent (blue dye) at same concentration for all ranges Stable and traceable to the National Institute of Standards and Technology, USA (NIST) 13
14 QC Kit Verification Plates Lot characterized plates Extensive testing of every plate lot Well dimensions traceable to the National Institute of Standards and Technology, USA (NIST) Bottom diameter (D) and taper angle ( ) are critical to volume calculations 2D barcode on plate boxes carries necessary information about performance, dimensions and lot number D 14
15 How it works - Overview of Dual-Dye Photometric Method Photometric measurement of liquid volume Two dyes measured at two wavelengths Ratiometric calculation of results per well Simultaneous measurement of accuracy & precision per tip Artel s patented dual-dye ratiometric photometry is included in ISO 8655: Part 7 and the measurement results are NIST-Traceable 15
16 Patented dual-dye Ratiometric technology from Artel The Tecan Infinite reader measures absorbance at 520nm and 730nm Based on the optical density (OD) measurements, the absolute volume of pipetted Range Solution is calculated This calculation factors in data gathered from the Calibration and Baseline steps 16
17 QC Kit Software for EVOware Fully integrated with Freedom EVOware, enabling seamless, step-by-step control of the entire workflow. A wizard controls pipetting, shaking, reading, data analysis and reporting, providing a straightforward pass/fail result. 17
18 QC Kit Reporting Is designed to meet quality and regulatory needs with the following reporting options: QC Kit report one page summary of the verification test results and a pass/fail analysis QC Kit extended data report for more detailed, systematic analysis of liquid handling verification results, to be used for troubleshooting and liquid handling optimization 18
19 QC Kit Report OVERALL QC REPORT TO SATISFY QUALITY AND REGULATORY NEEDS Consists of: User Information Instrument information Software version information Pass/Fail requirements Average test results for all channels The reports can be printed or exported to the PDF file format. The data can also be exported as CSVs for further analysis, such as trending or data driven troubleshooting. 19
20 QC Kit Extended Data Report THE AUTOMATICALLY GENERATED PRINTOUTS LISTS THE OVERALL ACCURACY AND PRECISION (CV) AND GIVES VOLUME INFORMATION PER TIP Report shows the overall outcome (pass or fail) Color coding shows results for each individual well at a glance Easy to identify if a tip deviates from the expected volume 20
21 Reporting Module PRINT ANY REPORT OR EXPORT TO.PDF OR.CSV FORMATS 21
22 Reliable Quality Control QUALITY CONTROL ENSURES THAT YOUR INSTRUMENTATION S PIPETTING RESULTS ARE CONSISTENT AND COMPARABLE Consistent Proves that your instrument is performing well over time, whether that is weekly, monthly or annually Comparable Shows that pipetting performance is similar to other pipetting devices Demonstrates that your Freedom EVO is performing as intended 22
23 23
24 DOE Basics Inputs Process Responses 24
25 Two Quantitative Responses Precision (percent CV) Trueness (percent systematic error) 25
26 26
27 Some Constant Inputs One liquid handling system One liquid type (aqueous) One syringe size One dispense method One elevation (barometric pressure) 27
28 Three Controlled Input Variables Temperature quantitative Relative Humidity - quantitative Volume Arrangements - categorical 28
29 Three Tip Sizes Configuration Code Tip Size Volume V V V V V V
30 Four Test Volumes Configuration Code Tip Size Volume V V V V V V
31 Nine Environments Temperature Three Levels 15, 22.5, 30 C Relative Humidity Three Levels 30, 55, 80 %RH 31
32 Nine Environments 32
33 Six Volumes at Each Environment V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 33
34 34 Plus Replicates V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6 V1 V2 V3 V4 V5 V6
35 Should We Randomize? 35
36 Fixed Navigation 36
37 Example of one QC Kit test result A B C D E F G H Precision Std Dev = 0.71 µl CV = 0.35% Trueness Mean Volume = µl Systematic Error = % 37
38 Precision 38
39 1000 µl tips delivering 200 µl 39
40 200 µl tips delivering 100 µl 40
41 200 µl tips delivering 25 µl 41
42 200 µl tips delivering 10 µl 42
43 50 µl tips delivering 25 µl 43
44 50 µl tips delivering 10 µl 44
45 Trueness 45
46 1000 µl tips delivering 200 µl 46
47 200 µl tips delivering 100 µl 47
48 200 µl tips delivering 25 µl 48
49 200 µl tips delivering 10 µl 49
50 50 µl tips delivering 25 µl 50
51 50 µl tips delivering 10 µl 51
52 What s going on? 52
53 Vapor Pressure and Temperature 15 C 22.5 C 30 C 53
54 Aspiration Step is Critical 54
55 Water Vapor Interaction - Psychrometric Chart Temperature 55
56 Calculate Evaporation Potential Temp P sat RH P vap P evap C kpa % kpa kpa
57 1000 µl Tip at 200 µl 57
58 1000 µl Tip at 200 µl 58
59 200 µl Tip at 100 µl 100 µl Delivery 59
60 200 µl Tip at 100 & 25 µl 100 µl Delivery 25 µl Delivery 60
61 200 µl Tip at 100, 25 and 10 µl 10 µl Delivery 100 µl Delivery 25 µl Delivery 61
62 50 µl Tip at 25 µl 62
63 50 µl Tip at 25 & 10 µl 25 µl Delivery 10 µl Delivery 63
64 25 µl from two different tips 200 µl Tip 64
65 25 µl - Smaller tip is less sensitive 200 µl Tip 50 µl Tip 65
66 10 µl from two different tips 200 µl Tip 66
67 10 µl: Smaller tip is less sensitive 200 µl Tip 50 µl Tip 67
68 Sensitivities in order Configuration Sensitivity Code Tip Size Volume per kpa V % V % V % V % V % V % 68
69 Conclusions 1) Environmental conditions impact any pipetting that incorporates air. 2) Precision was found to be insensitive to temperature and humidity over the testing range. 3) Trueness was found to depend on the environmental conditions a. Trueness should be optimization under the local environmental conditions b. A linear correlation was found between the trueness and the evaporation potential c. Fuller tips are less sensitive c. A stable environment will minimize impact on the business process 4) The QC Kit is unaffected by the environmental conditions. 69
70 Final Note The precision of the Tecan Freedom EVO was found to be insensitive to environmental conditions and performed within manufacturer s limits over the entire range tested in this study (15 C to 30 C and 30% to 80% relative humidity). Note: Tecan specifies the pipetting behavior of the Freedom EVO under controlled environment conditions (T = 20 C to 27 C; relative humidity at 25 C between 30% and 60%). Volumetric performance specifications apply to this controlled limit. For operation outside these limits meeting the manufacturer's specifications for volumetric performance is not guaranteed. 70
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