The core facility houses a variety of equipment for cloning, expression and purification of bacterially expressed recombinant proteins; this includes vectors for ligation-independent cloning (LIC), numerous strains of E. coli for expression of challenging constructs and a refrigerated incubator for large-scale production of temperature-sensitive proteins. For large-scale purification, the lab has a GE AKTA Purifier FPLC system equipped with a variety of columns for size exclusion, affinity and ion exchange chromatography.
Surface Plasmon Resonance
A SensíQ® Pioneer SPR system is available for characterizing protein-ligand interactions. The machine has three channels and includes an autosampler and two temperature-controlled slots for SBS-footprint deep-well blocks. This instrument offers a significant advantage over the more frequently used GE BiaCore models as it can derive kinetic parameters from single "OneStep" injections.
The core is equipped with a Rigaku MicroMax 007-HF X-ray generator and a Rigaku R-AXIS IV++ image plate detector. Cryo-equipment includes an Oxford Cobra cryostream, an inverted phi axis for easy retrieval of cryo-cooled crystals from the X-ray goniometer head and a SAM cassette for long-term storage of crystals under cryo-conditions. For experiments which may require a more intense X-ray beam, such as for high throughput data collection, or a tunable X-ray source, the facility also has rapid access to the SSRL synchrotron at Stanford University.
Two ARI robotic systems have recently been added to the core facility; a Gryphon for nano-liter liquid handing of SBS-footprint plates, and a Scorpion for custom liquid handling. The former is primarily used for setting up protein/DNA crystallization trials - using as little as 20 μL to screen 96 reagent conditions. The Scorpion is a more versatile robot which is principally used in optimization of crystal "hits." Two incubators set at 4°C and 18°C are dedicated for housing crystallization plates.
Additional Biophysical Instrumentation
In addition to the equipment mentioned above, the molecular therapeutics core facility includes GE MicroCal calorimeters for thermodynamic analysis of protein-ligand interactions through ITC. A Waters Breeze 2 HPLC equipped with a number of columns for small molecule and protein chromatography is also available for use.