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Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
Microplate Assays and High Throughput Screening HTPC
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Microplate Assays and High Throughput Screening HTPC

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pharmaceutical

pharmaceutical

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  • 1. M I C R O P L AT E A S S AY S : T H E K E Y P L AT F O R M F O R H I G H T H R O U G H P U T SCREENING IN BIOMEDICAL RESEARCH AND PHARMACOLOGICAL
  • 2. Microtiter Plate • develop in 1990s • 2:3 rectangle matrix with wells • 6, 24, 96, 384, 1536 wells • used for Enzyme-linked Immunosorbent Assay, High Throughput Screening, storage • Flat-shaped, U-shaped, V-shaped
  • 3. Well shape
  • 4. ELISA
  • 5. Microplate Reader taken from Synergy
  • 6. Reusing plate taken from © 2014 Kinesis Ltd. taken from Labotal Scientific Equipment (1997) LTD
  • 7. Materials • polyprolyene- for wide change in temperature environment • polycrabonate- disposable, • Cyclo-olefins
  • 8. High Throughput Screening an optimized, miniaturized assay format that enables the testing of > 100,000 chemically diverse compounds per day.
  • 9. Biotechnology Research Institute The Hong Kong University of Science and Technology • TCMs, including those for treatment of Alzheimer’s disease, neuro-protection, stroke, and sleep disorders
  • 10. taken from RNAi @ NIH Chemical Genomics Center
  • 11. History of Developing a Drug ! Drug discovery ! Initial characterisation ! Pre-clinical trials ! Regulatory approval sought to start trials in humans ! Clinical trials Phases I, II, III ! Submission of marketing/manufacturing authorization application to regulatory authorities ! Regulatory authorities review information and grant (or refuse) licences ! Product goes on sale ! Post-marketing surveillance ! Library of compounds ! In vitro screening: human/animal receptor/enzyme assay; reporter system ! Hits/lead ! Biochemical, tissue or animal model of function ! lead ! Animal model of therapeutic target ! ADME, formulation, acute toxicology
  • 12. High throughput screening for drug discovery high throughput screening (HTS) is number one tool for early-stage drug discovery ! HTS is process by which large numbers of compounds are rapidly tested for their ability to modify the properties of a selected biological target. Goal is to identify ‘hits’ or ‘leads’ - affect target in desired manner - active at fairly low concs (∴ more likely to show specificity) - new structure ! ! HTS = 50,000-100,000 cpds screenings per day!!!
  • 13. taken from Pete the Blogger
  • 14. ! Aim of screening is to find progressible hits, not to discover the lead molecule itself ! ! The majority of drug targets are a) G-protein coupled 7 TM receptors (est total 5000) b) nuclear receptors c) ion channels d) enzymes (est total >150) (est total 1000) (est total uncertain)
  • 15. Assays Definition - a test system in which biological activity can be detected • Aequorin assays • Colorimetric assays • Membrane potential assays • Intracellular calcium flux assays • Fluorometric assays • Luciferase reporter gene assays • Fluorescent and radiolabeled ligand binding assays
  • 16. Fluorescent Polarization Assay taken from GlycoForum
  • 17. Aequorin assay
  • 18. Luciferase Reporter Gene Assay from Thermo Scientific
  • 19. Virtual Screening • computational technique used in drug discovery to search libraries of small molecules in order to identify those structures which are most likely to bind to a drug target, typically a protein receptor or enzyme
  • 20. • Ligand based- logic based rules feature substructures and chemical properties of the candidate ligands • Structure based- virtually docking candidate ligands to the receptors
  • 21. The End

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