Based on in vitro assays, we performed a High Throughput Screening (HTS) to identify kinase inhibitors among 10,000 small chemical compounds. In this didactic paper, we describe step-by-step the approach to validate the hits as well as the major pitfalls encountered in the development of active molecules. We propose a decision tree that could be adapted to most in vitro HTS.

Reporter-based assays underlie many high-throughput screening (HTS) platforms, but most are limited to in vitro applications. Here, we report a simple whole-organism HTS method for quantifying changes in reporter intensity in individual zebrafish over time termed, Automated Reporter Quantification in vivo (ARQiv). ARQiv differs from current "high-content" (e.g., confocal imaging-based) whole-or...

Motivation: High-throughput screening (HTS) is an early-stage process in drug discovery which allows thousands of chemical compounds to be tested in a single study. We report a method for correcting HTS data prior to the hit selection process (i.e., selection of active compounds). The proposed correction minimizes the impact of systematic errors which may affect the hit selection in HTS. The in...

High-throughput screening (HTS) is an efficient technological tool for drug discovery in the modern pharmaceutical industry. It consists of testing thousands of chemical compounds per day to select active ones. This process has many drawbacks that may result in missing a potential drug candidate or in selecting inactive compounds. We describe and compare two statistical methods for correcting s...

  Photoproteins are excellent reporter systems because they don’t have virtually background signal. Aequorin is the most well-known photoprotein. Three improved engineered photoproteins photina, i-photina and c-photina, were also recently developed and optimized for generation of Ca2+ mobilization assays precisely. The total light emission is greater than aequorin and their reacti...

High throughput screening (HTS) involves using automated equipment to test a large number of samples against a defined molecular target to identify a reasonable number of active molecules in a timely fashion. Major factors which can influence priorities for the limited resources of the HTS group are projects, productivity, progress and people. The challenge to the HTS group is to provide excell...

In view of the essential role played by dosRS in the survival of Mycobacterium in the infected granuloma cells, dosRS transcriptional regulatory proteins were considered as a validated target for high throughput screening (HTS). However, the cost and time factor involved in screening large compound libraries are an important hurdle in identifying lead compounds. Therefore, the use of computatio...

The application of genome scale RNA interference (RNAi) relies on the development of high quality RNAi high throughput screening (HTS) assays. An important quality control (QC) characteristic in an HTS assay is how well the positive controls, samples, and negative controls can be separated from each other in the assay. Signal-to-noise ratio, signal-to-background ratio, Z-factor and Z’-factor ha...

Fragment screening is the process of identifying relatively simple, often weakly potent, bioactivemolecules. It is gainingwide acceptance as a successful hit-finding technique both in its own right and as a method of finding hit molecules when traditional high-throughput screening (HTS) methods fail. Fragment hits are typically highly ‘ligand efficient’, i.e. possess a high binding affinity per...

High throughput screening (HTS) has historically been used for drug discovery almost exclusively by the pharmaceutical industry. Due to a significant decrease in costs associated with establishing a high throughput facility and an exponential interest in discovering probes of development and disease associated biomolecules, HTS core facilities have become an integral part of most academic and n...