High throughput screening

Manish Kumar Mangalayatan university

1. High Throughput Screening (HTS) Submitted to: Mr. L.Ratnakar Singh Submitted by: Manish Kumar M.Pharm (Pharmacology) Department of Pharmacy , Institute of Bio-Medical Education & Research, Mangalayatan university Beswan, Aligarh

2. High Throughput Screening (HTS) Identification of one or more positive candidates extracted from a pool of possible candidates based on specific criteria.

3. Introduction High Throughput Screening (HTS) is a drug-discovery process widely used in the pharmaceutical industry. It leverages automation to quickly assay the biological or biochemical activity of a large number of drug-like compounds.

4. It is a useful for discovering ligands for receptors, enzymes, ion-channels or other pharmacological targets, or pharmacologically profiling a cellular or biochemical pathway of interest. Typically, HTS assays are performed in "automation-friendly" microtiter plates with a 96, 384 or 1536 well format.

5. Cell growth tests (cell-based assays or Phenotypic assays) Tissue response - targeted functional cell-based assay Enzyme test - biochemical test Assay Technology in HTS

6. Advantages of HTS High sensitivity of assay (single molecule detection) High speed of assay (automation) Minimization of assay ( microtiter plate assay) Low background signal Clear message (best: Yes/No answer)

7. Low complexity of assay (specific interaction) Reproducibility Fast data processing of results Acceptable costs !!!

8. Detection Methods in HTS: Spectroscopy Mass Spectrometry Chromatography Calorimetry X-ray diffraction Microscopy Radioactive methods

9. Spectroscopy in HTS: Fluorescence Spectroscopy Total internal reflection fluorescence (TIRF) Nuclear magnetic resonance (NMR) Absorption and luminescence sp. Fourier transformed infrared sp. (FTIR) Light scattering

10. Chromatography in HTS: Gas chromatography (GC) Thin layer chromatography Liquid chromatography (HPLC) Ion Exchange chromatography Reverse phase chromatography Hydrophobic interaction chromatography Affinity chromatography

11. Calorimetry in HTS: Isothermal titration Calorimetry (ITC) Differential scanning Calorimetry (DSC)

12. Microscopy in HTS: Scanning Tunnelling Microscopy Atomic Force Microscopy Confocal Microscopy

13. Uses of HTS: To screen for all kind of novel biological active compounds (libraries): Natural products Combinatorial Libraries (peptides, chemicals…) Biological libraries To screen Micro arrays such as: DNA chips RNA chips Protein chips