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
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