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7/31/2019 High-throughput Screening (Hts)
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HIGH-THROUGHPUT SCREENING (HTS)
- KARAN SAMYANI
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Without ability to screen libraries rapidly foractivity, there would be no combinatorialchemistry so that biologist are just as adeptat developing rapid HTS
HTS is very broad topic including enzymes,cells, organic cells, various tissue and whole
organs. Even sometimes whole animal too.
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HTS programs integrate
with. Target identification i.e genomics and
molecular group
Reagent preparation i.e protein expressionand purification of groups
Compound management i.e informationmanagement group
Assay development i.e biologist andpharmacologist involvement
High throughput library screening
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Formerly these activities were handled
separately but now it becoming more commonto integrate. By this way it will increasing thescreening efficiency
By using high density screening plate we canalso improve screening efficiency
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One of the first activity in developing
HTS assay is selecting the target. About500 target is currently being use. Amongthese- cell membrane receptor- makes
the largest group (45%) followed byEnzymes (28%).
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Concern about screening ?
HTS involve the screening compound that areapart of the corporate storehouse of thecompounds synthesized in the past or they may
be purchased from a vendor Such libraries consist of microtiter plate consist
of frozen or dried samples of compounds
The cost of complete screening may be over to
300000 USD. This type of screening is rather infrequently than
day to day screen.
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One can reduce the screening effort by poolingthe group of structure and running assays onmixture of compounds. By this we can alsoconserve reagents and biologic materials too.
A number of factors affect on screening like
solubility of compound , its ionizing andreacting capacity.
Even some compounds are structurally very
similar that may rise false positive hits this islikely to arise by pooling.
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To be effective, such compounds must dissolves
completely in the assay medium. Thats why itsvery common to add a small amount 1% ofDMSO.
The best concentration of compound to use issomewhat detectable. High con. Lead to morefalse positive results than that happens at lowconcentration.
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Method for detection
There are many ways by which we canmeasure the activity of assay.
These methods must be Reproducible,Accurate and High signal- to noise ratio
Two types of method are there
1)Non Radiometric
2) Radiometric
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Non radiometric method
It includes spectroscopy method
1) Absorbance
2) Fluorescence3) luminescence
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Radiometric method
It include
1) Scintillation proximity assay
2) filtrationthese assays include radioisotopes, so safe
storage and handling are of concern
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THANK YOU