Screening Support
鈥婽he HTCB can assist researchers with screening projects using a variety of assistance models. For occasional users the center can provide end to end support, starting with assay design and development, transitioning to full HTS campaigns, and concluding with data analysis and report generation.
鈥婱ore advanced users with HTS-ready systems can be assisted to translate these screens to our platform, select appropriate compound libraries, and process output data to prioritize screening hits.
鈥婩inally, users with extensive experience in HTS environments can obtain training to run the platform independently, reducing screening costs and permitting additional assay protocol optimization.
Experiments
The HTCB offers a suite of standard high-throughput screens to both internal and external users at very competitive rates. These include antimicrobial assays against a range of clinically relevant bacterial pathogens, pure protein assays, and high-content cytological profiling assays against a set of human cancer cell lines.
If required these assays can be run as fee-for-service experiments using owner-generated compound code numbers so that your proprietary data remains secure and private. Data is stored in our online data management system, permitting easy access from anywhere in the world, but limiting access to your selected research team. Users can access only the data in their own project, meaning that you and only you have permission to view, analyze and download your screening results.
BioMAP
AntiBiotic Mode of Action Profile (BioMAP) enables identification of compounds with potential anti-bacterial activity. BioMAP involves the use of high-throughput screening methodology to screen a panel of 19 diverse pathogenic bacterial strains, including both Gram negative and Gram positive strains, against compounds of interest. Subsequently, compounds with potential antibiotic activities are identified via calculation of percent inhibition values. As well, BioMAP based activity profiles of unknown compounds are clustered with activity profiles of known antibiotics, thereby enabling the categorization of unknown compounds with potential antibiotic activity according to antibiotic class. This assay is based on this protocol: https://doi.org/10.1016/j.chembiol.2012.09.014
MTT assay
The MTT assay determines the effect of a compound on cell viability and proliferation via assessment of mammalian cell metabolic activity. Metabolically active mammalian cells reduce tetrazolium salt (MTT) resulting in the production of purple formazan crystals. Subsequently the formazan crystals are solubilized, following which cell viability is assessed by measuring absorbance via a plate reader. A reduction in absorbance, compared to untreated controls, indicates reduced metabolic activity, which enables compounds that may affect cell viability and proliferation to be identified.
Cell Painting
Cell painting is a phenotypic assay which enables unbiased data associated with the physiological state of cells to be obtained via high-content microscopy. A set of five commercially available stains are utilized to assess the impact of compounds on six cellular compartments (endoplasmic reticulum, mitochondria, cytoskeleton, Golgi apparatus, nucleus, and nucleolus). During cell painting analysis, approximately 1000 measurements are extracted per image using Cell Profiler. These measurements are then clustered to provide an activity profile, or fingerprint, of a compound of interest. Subsequently, the fingerprints of unknown compounds can be clustered with the fingerprints of compounds with known biological activity, in order to identify potential cellular pathways that novel compounds of interest may act on. This assay is based in this protocol: https://doi.org/10.1038/nprot.2016.105