High Throughput

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High Throughput And High Content Screening
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What Is HTS ?
Biochemical target ? Test many compounds ? Compoun
ds with desired effect on target Hit ?
OR ? ? ?
Hit directly used as chemical probe to study
target
Hit provides structure for chemists to begin
developing a drug or other product
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HTS Technology
Robotics Miniaturization Sophisticated Assay
Chemistry Sophisticated Software and Database
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Evotec uHTS System

• Opera High Content Detector
• Platevision detector for fluorescence (also
  fluorescence polarization fluorescence
  resonance energy transfer), luminescence,
  absorbance
• 2 CO2 incubators
• 4 low volume pipetting systems for 384- and
  96-well plates (multidrops, Cy-Bi well, Cy-By
  drop)
• Plate washer
• Centrifuge
• Piercer
• Plate storage, barcode readers, lid handlers,
  transport systems

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platevision detector

• CCD camara
• Xenon lamp light source
• Zeiss optics
• Built in algorithm to correct for spherical lens
  aberration
• 4 filter sets
• Excitation filter
• Emission filter
• Dichroic filter

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Opera

• 4 lasers
• 1 xenon lamp
• 4 CCD cameras
• 96- and 384-well plates
• Acapella image analysis software

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Software Systems

• Genedata
• Assay Analyzer
• Condeseo
• Integrated with Opera Acapella image analysis
  software
• Accelyrs Accord
• HTS
• With Pipeline Pilot
• Ohio Third Frontier Computing Systems

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HTS
Output is QUALITY DATA
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Standard Screening Procedure

• 1o HTS ? hits
• Triplicate assay ? confirmed HTS hits
• 10 point dose-response assay ? EC50

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HTS Assay Examples

• Receptor-binding
• GPCR
• Enzyme
• protease, kinase, phosphatase, lipase, others
• Bacterial growth
• Cell-based reporter gene
• Cell growth, cell viability
• Cytochrome P450 inhibition
• Protein binding
• Protein production by cells
• ELISA

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HIGH-THROUGHPUT SCREENING
HIGH-THROUGHPUT SCREENING
Examples of Assays
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Results from Enzyme Inhibition Screen
11,500 compounds screened Normalized results
shown Each tile shows 1 plate of data
Right Color Scale for Normalization Distribution
of compound Activity
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Results from 1 plate in Enzyme Inhibition Screen
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High Content Screening With the Evotec Opera
96-, 384-, and 1536-well plates Up to 70
images per well in 384-well plate
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Internalisation Assay Example Image
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Internalisation Assay Example Image Analysis
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Internalisation Assay Example Opera External
Data output
Excel spreadsheet spots / cell cell
area nuclei area
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High Throughput High Content Screening Services

• Quality Data
• Experienced Scientists
• State-of-the Art Equipment

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Contacts

• Sandra Nelson
• High Throughput High Content Screening
• sandra.nelson_at_uc.edu
• 558-2372
• Ruben Papoian
• Drug Discovery
• ruben.papoian_at_uc.edu
• 558-0516
• Charles McOsker
• Business Development External Relations
• charles.mcosker_at_uc.edu
• 558-2569