high throughput and high content screening. what is hts ? biochemical target test many compounds ...

High ThroughputHigh Throughput

AndAnd

High ContentHigh Content

ScreeningScreening

What Is HTS ?

Biochemical target

Test many compounds

Compounds 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 drugor other product

HTS TechnologyHTS Technology

Robotics

Miniaturization

Sophisticated Assay Chemistry

Sophisticated Software and Database

Evotec uHTS SystemEvotec uHTS System

• Opera High Content Detector• Plate:vision 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

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

OperaOpera

4 lasers

1 xenon lamp

4 CCD cameras

96- and 384-well plates

Acapella image analysis software

Software SystemsSoftware Systems

Genedata• Assay Analyzer• Condeseo• Integrated with Opera Acapella

image analysis software

Accelyrs Accord• HTS • With Pipeline Pilot

Ohio Third Frontier Computing Systems

HTSHTS

Output isOutput is

QUALITY DATAQUALITY DATA

Standard Screening ProcedureStandard Screening Procedure

1o HTS hits

Triplicate assay confirmed HTS hits

10 point dose-response assay EC50

HTS Assay ExamplesHTS 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

HIGH-THROUGHPUT SCREENING

Examples

of Assays:

HIGH-THROUGHPUT SCREENING

Results from Enzyme Inhibition ScreenResults from Enzyme Inhibition Screen

11,500 compounds screenedNormalized results shownEach tile shows 1 plate of data

Right:Color Scale for NormalizationDistribution 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

spots per cell

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

1 3 5 7 9 11 13 15 17 19 21 23

spots per cell

Internalisation Assay Example

Opera External Data output

Excel spreadsheet

spots / cell

cell area

nuclei area

High Throughput & High Content High Throughput & High Content Screening ServicesScreening Services

Quality Data

Experienced Scientists

State-of-the Art Equipment

ContactsContacts

Sandra Nelson• High Throughput & High Content Screening• sandra.nelson@uc.edu• 558-2372

Ruben Papoian• Drug Discovery• ruben.papoian@uc.edu• 558-0516

Charles McOsker• Business Development & External Relations• charles.mcosker@uc.edu• 558-2569