Workflows requiring high-throughput screening

The primary goal of high-throughput screening is to streamline the drug discovery process by rapidly analyzing large numbers of potential biological modulators to identify those with the desired effect on a specific target. HTS is typically used in the following workflows:

• Drug Discovery: Identifying potential drug candidates that interact with specific biological targets.
• Functional genomics: Understanding gene function and identifying genes that play a role in specific pathways.
• Toxicology: Evaluating the potential toxicity of various compounds.
• Pathway analysis: Identify key players in specific cellular pathways.

Overview of the high-throughput screening process

Step 1: Sample and compound library assembly

This step requires the selection of biologically relevant targets, typically proteins or cellular pathways known to play a role in disease or other relevant processes. At the same time, various compounds are created or sourced. These compounds, which range in number from thousands to millions, constitute “compound libraries” that will be screened for potential activity against the target.

Step 2: Develop assays compatible with high-throughput

Central to the HTS process is the design and application of effective detection methods. This is no ordinary test; it is carefully tailored to measure the impact of the compounds in the library on a specified biological target. To be effective in HTS environments, detection methods must be ultra-sensitive to the smallest changes and have the flexibility to manage large numbers of samples. Additionally, the transition to automation requires assays to be optimized for devices such as plate readers, which often means assays need to be customized for formats such as 96-well or 384-well plates. In addition, it is critical to ensure that the assay is stable and reproducible across a large number of samples.

• Cell-based assays: Here, live cells occupy the central position. Test compounds within these cells to determine their effects, whether on cell health, proliferation, or unique cellular responses.
• Biochemical tests: Further scaling up, these experiments use purified proteins or specific biomolecules to determine the effects of compounds. Usually, this can be enzyme kinetics exploration or subtle binding experiments.
• Phenotypic determination: From a broader perspective, these experiments delve into the entire field of living organisms. By introducing compounds into entities such as zebrafish or Korean fish, researchers can monitor any significant changes in phenotype.

Step 3: Automate infrastructure setup

Due to the large number of samples and compounds, manual processing is impractical. Therefore, a specialized pipetting workstation is required to automate the process. These workstations can handle tasks ranging from precise dispensing of sample and compound amounts to transferring liquids between microplates. The entire unit is designed to minimize errors, increase speed, and ensure consistent processing of each sample.

Step 4: Data collection and analysis

When compounds are screened using detection methods, a large amount of data is generated. Automated plate readers, imaging systems, or other detection methods can be used to collect this data. Specialized software then processes this information and analyzes it to find compounds that show the desired effect, known as "hits." This step is crucial as it sets the stage for further research and optimization based on these promising compounds.