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Structural Biology Services: Protein Crystallization, X-ray Crystallography and NMR Spectroscopy

OVERVIEW
Protein crystallization and structure determination by X-ray crystallography as well as nuclear magnetic resonance (NMR) spectroscopy are the main methods for high resolution protein structure determination. The main components of the structural biology contract services at SARomics Biostructures include:

  • high-throughput protein expression and purification,
  • crystallization of proteins and ligand protein complexes and X-ray structure determination
  • antigen-antibody complex structures & biosimilars characterization
  • protein optimization
  • protein NMR spectroscopy
  • fragment screening and fragment-based drug discovery
With proven expertise in structure determination and analysis of biological macromolecules, we add to our customers’ projects clear insights into the mode of binding and the mechanisms of interactions in ligand-protein and antigen-antibody complexes. In addition, our structure-based drug discovery services are based on fragment library screening using our proprietary weak affinity chromatography technology (WAC™), hit expansion and optimization guided by protein-ligand complex structures. The following pages describe details of the technology platform and are design to help our customers in the preparation for the experiments:

Crystallization and crystallography (fee-for-service crystallography)
• Protein NMR spectroscopy
Computational chemistry & drug discovery


SARomics’ structural biology platform include all major techniques required for successful protein structure determination. Following cloning, expression and purification of the protein (gene-to-structure services), and before the start of an X-ray crystallography or NMR spectroscopy project, we perform a thorough biophysical characterization of the protein in solution - both for successful crystallization and for NMR structure studies we need monodisperse solutions, which do not contain any aggregated or denatured material. The protein also needs to be stable within a certain range of pH, ionic strength and temperature (these values will be varied when crystallization screens are run). It should always be kept in mind that a proper characterization of the protein solution is crucial for the success of a structure determination project!

Several biophysical techniques are routinely used at SARomics Biostructures for protein characterization. Among those is:

  • NMR spectroscopy,
  • dynamic light scattering (DLS),
  • CD spectroscopy
  • differential scanning fluorimetry (DSF, similar to thermal-shift assay).
  • microscale thermophoresis (MST)

After crystallization, high resolution X-ray diffraction data are collected and the crystal structure is determined by interpreting the calculated electron density map.