The TVS regularly performs a wide range of in vitro protein-protein (fluorescene polarization competition assays, FRET, TR-FRET, AlphaScreens, AlphaLISA) and protein-ligand (lanthanide metal competition assay) assays and cell based assays include mammalian cell viability, chemiluminescence gene reporter assays, yeast and bacterial growth, and cell based ELISAs for screening small drug like compounds. The center currently has 60,781 small drug like compounds available for screening that our maintained by the TVS personnel to assure the highest purity and maintain control of compound inventory using ChemBioFinder Software (ChembridgeSoft). In addition, the TVS program has access to a Bruker 800 MHz NMR equipped with a 60-sample auto-loader to run NMR based screens in conjunction with the Structural Biology (SB) that allows for direct binding of ligands to proteins via NMR experiments. The NMR is particular useful in fragment-based lead discovery a new lead discovery approach in which much lower molecular weight (100-300Da) compounds are screened relative to traditional drug screening campaigns. Fragment-based hits are typically weak inhibitors and therefore need to be screened at higher concentration using very sensitive biophysical detection techniques such as protein crystallography and NMR as the primary screening techniques. While theses fragments are simpler, less functionalized compounds with correspondingly lower affinity, the final goal is the chemical crosslinking of multiple fragments increasing their affinity and specificity so the new compound is suitable for optimization into clinical candidates with good drug-like properties.
The TVS group has biosafety cabinets, CO2 incubators, an inverted phase microscope, low speed centrifuges, and dedicated liquid nitrogen storage freezers. All laboratories have refrigerators, freezers, house vacuum, hood ventilation, and natural gas hook-ups. Autoclave, dishwashing, and pure water facilities are located adjacent to the laboratory space. Office suites (PI, postdocs, students and staff) are clustered together and located adjacent to the laboratory space. All offices are equipped with desktop computers and broadband internet access is freely available throughout the site (wired and wireless). The IBBR complex has a business office to assist with purchasing/receiving as well as on-site information technology support and facilities support.
TVS has are two automated liquid handling stations. The Biomek FX with a 96-well head capable of rapid liquid transfers in 96, 384, and 1536-well plates and large reservoirs and moving plates and pipette tip boxes within the 23 deck positions. The Biomek NXp is equipped with an 8-span head that has 8 independent pipette tips and 11 positions on the deck. While slower at some task than the Biomek FX it is capable of manipulations and reformatting in vials, reservoirs, tubes, and multitude of assay plates including 6, 12, 24, 48, 96, and 384-well plates. The three BMG multimode plate readers equipped with autoloaders (1) BMG Optima POLARstar (2) BMG PHERAstar Plus, and (3) BMG PHERAstar FS, that together have the ability to read assays in 96, 384, 1536, and even 3456-well plates in several detection modes including fluorescence intensity, fluorescence polarization, fluorescence resonance energy transfer (FRET), time resolved fluorescence (TR), luminescence, UV/Vis absorbance spectra.
The TVS has 60,781 small drug like compounds available:
- The ChemDiv 40K Collection is composed of 40,000 diverse chemical compounds purchased from Chemical Diversity Labs (ChemDiv) in a 384-well format, each at 10 mM in 100% DMSO.
- The Maybridge HitFinder Collection is a collection of 14,400, drug like compounds from Maybridge special packaged as a dry film with 0.1 cm of each compound in a 384-well plate format that are then typically suspended in 100% DMSO at 10 mM.
- The MicroSource Spectrum Collection is made up of 2,000 compounds, 50% drugs, 30% natural products, and 20% other bioactive components available in a 96-well format, each at 10 mM in 100% DMSO.
- The SBi Collection is a collection of 4,381 compounds collected from several commercial sources plus 67 compounds synthesized in house (by Andrew Coop Lab).
The TVS has just purchased the Maybridge Ro3 Fragment a collection of 1,000 compounds meeting the Rule-of-3 Fragment Library offers keys features necessary and available in many fragment libraries while guaranteeing solubility of the compounds @ 200 mM in DMSO and having chemically linkable analogs available for future linking of fragments into higher affinity compounds.