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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