High Throughput Screening (HTS) is a drug discovery process that uses automation to quickly assay a large number of compounds against biological or biochemical targets to identify potential drug candidates. Key aspects of HTS include testing compounds in microtiter plates with 96, 384, or 1536 wells using assays like cell-based, enzyme, or tissue response tests. HTS allows for high speed, sensitivity, and reproducibility in screening large libraries of compounds cost effectively. Detection methods used in HTS include spectroscopy, chromatography, calorimetry, and microscopy.

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