General Description
The Core Facility for Crystallography and Biophysics offers services from small molecule crystallography, trough in vitro screening and biochemical and biophysical characterization to the structural biology of proteins and their complexes with ligands, including:
- Computational theoretical studies on small molecules and their complexes with cellular targets
- Small molecule crystallization process
- Small molecule properties and their polymorph analysis
- Small molecules and protein X-ray crystallography
- Protein engineering and expression
- Biochemical and biophysical studies on ligand-target analysis.
Most services are optional (used as a stand-alone) and the sample may enter and exit CFCB pipeline (CHEM or BIO) at any point, or as a or all-in-one service. The appropriate reports are modular for that purpose.
Service Features
Ligand and Protein Binding Assays
Method or Service
MicroScale Thermophoresis (MST)
- Ligand or protein partner binding
Thermo Fluor Assay
- Buffer and additive selection
- Ligand binding assay
Enzymatic activity assays and readouts
- Protein activity or concentration
- Ligand binding
SEC-MALS
- Protein partner binding
- Protein-detergent complexes
Report to Customer
- Best protein construct
- Suggested buffer for sample stabilization
- Suggested ligand choice
- Suggested detergent
- Ligand or protein partner Binding (Kd)
- Protein activity (Vmax, Km, kcat…)
Quality Check and Optimization
Method or Service
SDS-PAGE analysis (1 D or 2 D)
Spectrophotometer plate reader
Enzymatic activity assays and readouts
- Dynamic Light Scattering (DLS)
Report to Customer
- Concentration
- Purity
- Homogeneity
SEC-MALS
- Aggregation assay
- Oligomerization state assay
- Molecular weight estimate
- Homogeneity
- Complex formation
Crystallization Trials
Method or Service
Initial screening and optimization
- JCSG+ Eco, Wizard I&II, Wizard III&IV, Midas, Morpheus, ShotGun(SG1), PACTPremier
- Custom or commercial condition sets
- Hanging/sitting drop (2 or 3 drops), sandwich plates, LCP
- Incubation at different temperatures
- Additive screens
- Seeding
- Soaking with heavy atoms for phasing
Customer-provided compound library
- Soaking or co-crystallization
Visualization and crystallization drop scoring
Report to Customer
- Crystallization conditions report
- Buffer composition
- Crystal morphology (including photographs)
- Temperature
- Additives
Diffraction and Data Collection
Method or Service
In-house Diffraction Data Collection
- Fishing, freezing, mounting
- Testing crystal diffraction quality
- Dataset collection for suitable crystals
External Synchrotron Trip
- Shipping and measurement, phasing
Report to Customer
- Crystals X-ray diffraction quality
- All or preliminary diffraction images and statistics
- Full diffraction dataset if collected successfully
Model Building
Method or Service
Structure solving and model building
- Crystallographic computing
Report to Customer
- Final model file and statistics
- Model deposition (PDB) if not confidential
Sample and Data Storage
- Samples, crystals or data can be handed over
