Manish Kumar Mangalayatan university

1. High Throughput Screening
(HTS)
Submitted to: Submitted by:
Mr. L.Ratnakar Singh 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. Assay Technology in HTS
 Cell growth tests (cell-based assays or
Phenotypic assays)
 Tissue response - targeted functional cell-
based assay
 Enzyme test - biochemical test

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