We have established multiple independent, parallel robotic systems for HTS/μHTS/iHTS and HCS operations that are capable of handling 96-, 384-, and 1536-well plate formats. These systems accommodate a variety of assay formats and are particularly well-suited for conventional and multiplex protein-protein interactions and phenotypic screens.
- HTS/HCS system I: The first system features a central vertical robotic system with three outpost readers: (i) EnVision multimode reader (HTS in 96/384/1536 well format), (ii) FlexStation II agonist-injectable, 384-well fluorescence reader, and (iii) ImageXpress (HCS in 96/384-well format). This system is integrated with a cell hotel, plate stacker, and various liquid handlers equipped with pin tools for low-volume (nL) transfer. All of these components are in an enclosed environment, facilitating live-cell HCS screens under aseptic conditions.
- HTS/HCS system II: The second system features the Twister II robot integrated with (i) an EnVision multimode reader (HTS in 96/384/1536-well format) and (ii) ImageXpress (HCS in 96/384/1536 well format) supported by the Sciclone liquid handling workstation.
- System III: Corning’s Epic system for label-free molecular interaction screens provides a third platform for hit identification and confirmation by direct detection of protein-compound binding and cell-based screens under physiological conditions.
To enhance the productivity of the HTS operation and the quality of hit discovery especially for multiprotein interactions, we have developed a number of multiplexed HTS strategies that we collectively term iHTS (information-rich HTS) to distinguish this format from HCS or conventional HTS. The iHTS format integrates multiple parameters in one well for protein-protein interaction modulator screens with the recognition that protein interactions often occur in a multiprotein complex and often with a selective member of the target protein family.
- HTS, biochemical and cell-based high-throughput screening in 384-well plate format
- μHTS,biochemical and cell-based ultra high-throughput screening in 1536-well plate format
- HCS, cell and organism-based, high content screening in 384/1536-well plate format
- iHTS, information-rich HTS in 384/1536-well plate format
- Biosensor-based label-free cell pathway screen in 384-well plate format
Agilent 6100 Series Quadrupole LC/MS is used for hit compound screening and molecular weight confirmation. The HTS team also executes quality control procedures (LC-MS, NMR, elemental analysis) on each confirmed hit that passes secondary screens to ensure the transfer of high quality hit compounds to the next phase of the project. To prevent unnecessary efforts around an impure compound, the structure and purity of hit compounds are confirmed using LC/MS, NMR, and elemental analysis when appropriate.
The CambridgeSoft Enterprise software serves as the backbone of the informatics environment at the center. It provides an integrated method of analyzing HTS/HCS assay results. Tools such as Inventory Loader, Inventory Manager, BioAssay HTS, and BioSAR are used for consolidating assay results into a central database for analysis and mining. The CambridgeSoft system consists of a CambridgeSoft Web Server that contains the Inventory Manager and BioSAR tools, an Oracle database, and a proxy server that provides the network connectivity of tools such as Inventory Loader and BioAssay HTS.
Example usages include:
- BioAssay Enterprise – data reduction, data management
- BioSAR Enterprise – data presentation, result query
- Registration Enterprise – compound library management
- Inventory Enterprise – compound library management, plate management
- ChemFinder – Sd file preprocessing, report preparation
- ChemDraw – manuscript preparation, chemical query definition
- ECBDC Labview (custom in-house project management software)
ECBDC supplements HTS/HCS capabilities and expertise with predictive calculations including 2D clustering, protein-ligand molecular modeling, receptor and ligand-based virtual screening, and ADME/Tox calculations. Information derived from these calculations is used to aid medicinal chemistry efforts to choose a lead from the myriad HTS hits and guide synthetic optimization of the selected lead compounds. The molecular modeling software tools used include LeadScope, Pipeline Pilot, Glide, and QikPro.