BMA Laboratories

1
Introduction
Jon Curtis GSK - ATG Systems Development The
Use of Adaptive Focused Acoustic Technology to
Improve uHTS Assay Performance and Compound
Dissolution
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Introduction

• Technology Overview
• Instrumentation
• Project areas
• Summary

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Technology overview
4
Technology overview treatments
Varying duty cycle/intensity/cycles per burst
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Technology
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Technology overview Video carbon nano tubes
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Technology overview Video
Yttrium bead in suspension
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Covaris Instrumentation E200 / S2

• Developed out of Lithotripsy-kidney stone
  disruption Ultrasound imaging
• Highly developed not a concept or prototype
• Over 200 units sold into DMPK ADMEtox metabolite
  RNA extraction

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Application areas Compound Management
Deliverables ATG

• Objective maintain compound concentration from
• HTC CM uHTS (XC50 SAR)
• Compound solubility issues caused by
• Initial dissolution - glues gums etc
• Precipitation - water uptake freeze thaw

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Application area Compound Management - video
Capable of dissolution in 20s that previously
would take 12 hours
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Application area Compound Management - video
Thawing and mixing of 2D tube
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Application area Compound Management - solution

• Solution
• Acoustic dissolution Primary secondary stage.
• Mixing/thawing in 4ml vials, 2D tubes and storage
  plates
• Rapid dissolution ?
• Automatable ?
• Compounds to thermodynamic equilibrium ?

No compound degradation observed lcms data
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C2000 automated acoustic dissolution
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Application area uHTS SCP AD
Objective Improve assay quality/data Solution
Covaris non contact acoustic mixing in low volume
assays in 384 1536 Preliminary Aims Improve
Z Reduce CV Re-solublising compound in
aqueous Accelerate assays
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Application areas uHTS - 1536 plate mixing video
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Application areas uHTS data
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Data
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Application areas uHTS preliminary conclusion
Impact 33 increase in Z of HTRF cAMP assay 9
increase in Z HTRF kinase assay 30 reduction in
assay time to reach equilibrium/maximal
Resulting in Increased signal window Increase
compound dissolution Faster assays Higher quality
screens Lower false positives Higher real
positives
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Application areas uHTS Membrane preps

• Membrane assays are often highly viscous making
  it very difficult to aspirate and dispense
  resulting in a poor assay Z.
• Caused by a combination of protein aggregation
  and high molecular weight genomic DNA.
• Requiring syringe treatment

• Conclusion
• 10-20 second Acoustic treatment
• Improve the preparation homogeneity
• EC50 assay equilibrium is attained after 1 hour
• Signal to noise increase 100
• Improved assay quality (Z)
• Recover failed assays.
• Simplify initial membrane prep itself

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Application area High Throughput Chemistry ATG
deliverables
Objective Automated dissolution of dried
compounds in methanol Solution Acoustic
mixing in low volume assays in 48 well CleVap
plates C200 Achievements Rapid dissolution
? Automatable ? Compounds to thermodynamic
equilibrium ?
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GSK Application areas

• High-throughput screening
• Compound Dissolution and Resolution
• Cell Lysis mammalian, Insect, E-coli, plant
• Continuous flow processing
• High Throughput Chemistry
• Drug Metabolism and Pharmacokinetics
• RNA extraction - Homogenisation of biological
  tissue
• Tissue homogenisation for Proteomic studies

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The end questions

• Many thanks to
• Zoë Blaxill CASS
• Jim Chan CM
• Ken Murray CM
• Karen Dobbs CM
• Todd Graybill HTC
• Suzanne Baddeley AD
• Liz Clark SCP
• Jim Laugharn Covaris
• Phil Robinson KBioscience