High Throughput And High Content Screening

What Is HTS ?

Biochemical target  Test many compounds  Compounds with desired effect on target =

Hit

  Hit directly used as chemical probe to study target OR   Hit provides structure for chemists to begin developing a drug or other product

HTS Technology

Robotics Miniaturization Sophisticated Assay Chemistry Sophisticated Software and Database

Evotec uHTS System

• • • • • • • • Opera High Content Detector Plate:vision detector for fluorescence (also fluorescence polarization & fluorescence resonance energy transfer), luminescence, absorbance 2 CO 2 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

plate::vision 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

Opera

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

Software Systems

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

HTS

Output is QUALITY DATA

Standard Screening Procedure

1 o HTS  hits Triplicate assay  confirmed HTS hits 10 point dose-response assay  EC 50

HTS Assay Examples

• • • • • • • • • Receptor-binding • GPCR Enzyme • protease, kinase, phosphatase, lipase, others Bacterial growth Cell-based reporter gene Cell growth, cell viability Cytochrome P 450 Protein binding inhibition Protein production by cells ELISA

Examples of Assays:

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

Results from 1 plate in Enzyme Inhibition Screen

High Content Screening With the Evotec Opera

96-, 384-, and 1536-well plates Up to ~ 70 images per well in 384-well plate

Internalisation Assay Example

Image

Internalisation Assay Example

Image Analysis

Internalisation Assay Example

Opera External Data output

Excel spreadsheet spots / cell cell area nuclei area spots per cell 0.7

0.6

0.5

0.4

0.3

0.2

0.1

0 1 3 5 7 9 11 13 15 17 19 21 23 spots per cell

High Throughput & High Content Screening Services

Quality Data Experienced Scientists State-of-the Art Equipment

Contacts

Sandra Nelson • • • High Throughput & High Content Screening [email protected]

558-2372 Ruben Papoian • • • Drug Discovery [email protected]

558-0516 Charles McOsker • • • Business Development & External Relations [email protected]

558-2569