HIGH-THROUGHPUT SCREENING (HTS)

- KARAN SAMYANI

 Without ability to screen libraries rapidly for

activity, there would be no combinatorial chemistry so that biologist are just as adept at developing rapid HTS HTS is very broad topic including enzymes, cells, organic cells, various tissue and whole organs. Even sometimes whole animal too.

HTS programs integrate with.

Target identification i.e genomics and

molecular group Reagent preparation i.e protein expression and purification of groups Compound management i.e information management group Assay development i.e biologist and pharmacologist involvement High throughput library screening

 Formerly these activities were handled

separately but now it becoming more common to integrate. By this way it will increasing the screening efficiency By using high density screening plate we can also improve screening efficiency

 One of the first activity in developing

HTS assay is selecting the target. About 500 target is currently being use. Among these- cell membrane receptor- makes the largest group (45%) followed by Enzymes (28%).

Concern about screening ?

HTS involve the screening compound that are apart of the corporate storehouse of the

compounds 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.

 One can reduce the screening effort by pooling the group of structure and running assays on mixture of compounds. By this we can also

conserve reagents and biologic materials too. A number of factors affect on screening like solubility of compound , its ionizing and reacting capacity. Even some compounds are structurally very similar that may rise false positive hits this is likely to arise by pooling.

 To be effective, such compounds must dissolves

completely in the assay medium. Thats why its very common to add a small amount 1% of DMSO. The best concentration of compound to use is somewhat detectable. High con. Lead to more false positive results than that happens at low concentration.

Method for detection

There are many ways by which we can

1) 2)

measure the activity of assay. These methods must be Reproducible, Accurate and High signal- to noise ratio Two types of method are there Non Radiometric Radiometric

Non radiometric method

It includes spectroscopy method 1) Absorbance 2) Fluorescence 3) luminescence

Radiometric method
It include 1) Scintillation proximity assay 2) filtration

these assays include radioisotopes, so safe storage and handling are of concern

THANK YOU