high throughput and high content screening. what is hts ? biochemical target test many compounds ...
TRANSCRIPT
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• [email protected]• 558-2372
Ruben Papoian• Drug Discovery• [email protected]• 558-0516
Charles McOsker• Business Development & External Relations• [email protected]• 558-2569