1. High Throughput And High Content Screening

2. 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 drug or other product

3. HTS Technology Robotics Miniaturization Sophisticated Assay Chemistry Sophisticated Software and Database

5. 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

6. 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

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

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

9. HTS Output is QUALITY DATA

10. Standard Screening Procedure 1 o HTS  hits Triplicate assay  confirmed HTS hits 10 point dose-response assay  EC 50

11. 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 inhibition Protein binding Protein production by cells ELISA

12. HIGH-THROUGHPUT SCREENING Examples of Assays: HIGH-THROUGHPUT SCREENING

13. 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

14. Results from 1 plate in Enzyme Inhibition Screen

15. High Content Screening With the Evotec Opera 96-, 384-, and 1536-well plates Up to ~ 70 images per well in 384-well plate

16. Internalisation Assay Example Image

17. Internalisation Assay Example Image Analysis

18. Internalisation Assay Example Opera External Data output Excel spreadsheet spots / cell cell area nuclei area

19. High Throughput & High Content Screening Services Quality Data Experienced Scientists State-of-the Art Equipment

20. Contacts 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