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All services at CMS are available both to unexperienced users and to experienced users (that do not require the assistance of the scientist in charge).

 

Biophysical techniques

 

  • Surface Plasmon Resonance (SPR) characterization. The service can be provided both with an SPR chip, or with the user bringing his/her own chip for use with the Biorad instrument;
  • Prometheus DSF assay, for the study of protein unfolding and aggregation detection;
  • Monolith NT.115 thermophoresis characterization;
  • Monolith NT.LabelFree thermophoresis characterization;
  • Isothermal titration calorimetry on the Microcal iTC200 instrument, label-free characterization of an interaction system;
  • Differential scanning calorimetry on the Microcal VP-DSC instrument. Determination of stability of biomolecular systems (both intramolecular and intermolecular), including that of micellar systems;
  • UV/visible precision spectroscopy, either at a single temperature or using a 5 – 95 ºC temperature ramp;
  • Dynamic light scattering measurements using either the Nano ZS90 apparatus or in-drop DLS with the Spectrolight600 apparatus;
  • Measurement of circular dichroism spectra and of absorbance on the Chirascan Plus CD spectrometer, to determine the type of secondary structure present in protein samples as well as the conformation of nucleic acids (DNA and RNA).

 

 

Crystallisation of proteins and nucleic acids/Diffraction techniques

 

  • Robotic setup of 96-well crystallisation plates, for screening of crystallisation conditions (also for “routine crystal production”)  -  for proteins, nucleic acids, complexes of biological macromolecules;
  • Manual setup of crystallisation plates, for screening, crystal optimization or “routine crystal production”, using different plate formats. Crystallisation can be setup in a cold room, at room temperature or in a warm room (26°C), all rooms being equipped with dedicated binoculars for plate setup and inspection;
  • Idem, under an inert atmosphere;
  • Automated monitoring of crystallisation in the Formulatrix crystal hotel. Lighting using visible light, polarized light and UV light. Remote access to crystallisation droplet images, and automated preliminary evaluation;
  • Crystal handling and preparation for diffraction experiments, including crystal fishing from droplets, cryo-protection, cryo-cooling (vitrification in liquid N2) and possibly long term storage in specialized Dewars containing liquid N2;
  • Idem, either in oxygen-free conditions or with the preparation of Xe derivatives, also possibly with long-term storage in specialized Dewars;
  • Supply of mounted cryo-loops;
  • In-situ (in the crystallisation plates) testing of crystal diffraction using the ISX stage;
  • Testing of diffraction using mounted crystals and / or measurement of X-ray diffraction data;
  • Diffraction data processing, providing a data file (such as an MTZ file);
  • Assistance/advice to solve a 3D structure (including a full 3D structure determination service on request);
  • Measurement of an X-ray diffraction data set at a synchrotron radiation source.

 

 

Structural mass spectrometry

 

  • Precise molecular weight determination by ultra-high resolution FT-ICR mass spectrometer with sequence confirmation by Top-down approach using different fragmentation techniques (collision induced dissociation, electron transfer/capture dissociation);
  • Peptide mass fingerprinting – identification of proteins from gel or solution including larger protein mixtures by using MALDI or ESI;
  • Characterization of posttranslational modifications such as phosphorylation, glycosylation or disulphide bonds;
  • Structural mass spectrometry: limited proteolysis, hydrogen/deuterium exchange, chemical cross-linking, covalent labelling;
  • HPLC separation of peptides, proteins and small molecules (metabolites) coupled with mass spectrometric detection by FT-ICR;
  • Processing and interpretation of acquired mass spectrometric data.