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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
HIGH THROUGHPUT SCREENING (HTS)

  Identification of one or more positive candidates
     extracted from a pool of possible candidates
              based on specific criteria.
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.
 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.
Assay Technology in HTS

 Cell   growth tests (cell-based assays or
 Phenotypic assays)
 Tissue   response - targeted functional cell-
 based assay
 Enzyme     test - biochemical test
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)
 Low    complexity of assay (specific interaction)

 Reproducibility


 Fast   data processing of results

 Acceptable    costs !!!
DETECTION METHODS IN HTS:
• Spectroscopy
• Mass Spectrometry
• Chromatography
• Calorimetry
• X-ray diffraction
• Microscopy
• Radioactive methods
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
CHROMATOGRAPHY IN HTS:
• Gas chromatography (GC)
• Thin layer chromatography
• Liquid chromatography (HPLC)
• Ion Exchange chromatography
• Reverse phase chromatography
• Hydrophobic interaction chromatography
• Affinity chromatography
CALORIMETRY IN HTS:

• Isothermal titration Calorimetry (ITC)
• Differential scanning Calorimetry (DSC)
MICROSCOPY IN HTS:

• Scanning Tunnelling Microscopy

• Atomic Force Microscopy

• Confocal Microscopy
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
High throughput screening

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High throughput screening

  • 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