BIO Pipeline for Biomacromolecules

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
  • 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

  • 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

Sample Preparation