I2BC / Plateforme de mesures d'Interactions Macromoléculaires (PIM)
Technological platform
The I2BC's Macromolecular Interactions Measurement Platform (MIP) offers a wide range of equipment and expertise in biochemistry and biophysics for quality control measurements and functional analysis of protein samples: microcalorimetry (ITC: Isothermal Titration Calorimetry and DSC: Differential Scanning Calorimetry), analytical ultracentrifugation (UCA), MST (Microscale Thermophoresis), SEC-RALS-LALS, BLI (Biolayer Interferometry), switchSENSE and more recently FIDA (Flow Induced Dispersion Analysis).
The services offered by the platform include: 1) Biophysical characterisation of proteins or biological complexes (SEC-RALS-LALS, UCA and FIDA) - 2) Analysis of biological interactions (ITC, BLI, switchSENSE, MST and FIDA) - 3) Analysis of the stability of macromolecules and biological complexes, in particular as an aid to formulation (DSC, DSF and TSA).
The equipment and services are available to all Université Paris-Saclay laboratories, other academic laboratories and industry.
For more information about these services: plateforme-pim@i2bc.paris-saclay.fr.
This platform is part of the I2BC Structural Biology Platforms cluster, which includes: i) the Crystallisation, NMR, CryoEM, Macromolecular Interaction Measurements and alphaRep platforms, and ii) the technical platforms for Soluble or Membrane Protein Expression in Yeast and Protein Expression in Insect Cells.
- Mrs Magali AUMONT-NICAISE
- Mrs Stéphanie MARSIN
- Mrs Magali NOIRAY
- Mr Stéphane PLANCQUEEL
Innovation themes
Lab of attachment
I2BC - UMR9198 avenue de la terrasse Bâtiment 24
91190 Gif sur yvette
Technologies utilisées
1) Biophysical characterisation of proteins or biological complexes:
- SEC-RALS-LALS (OMNISEC - Malvern): determination of the molar mass and stoichiometry of protein complexes, protein/detergent, protein/nucleic acid, protein/glycosylation.
- FIDA (Fida Biosystems): FIDA technology uses fluorescence detection of Taylor Diffusion Dispersed (TDA) systems in capillaries. By measuring the diffusion coefficients of species in solution from taylorgrams, the size of biomolecules can be calculated, accurately determining their hydrodynamic radius (Rh). It is then possible to characterise biomolecular interactions as a function of the size of the complexes formed.
- UCA, Optima XL-I and Fluorescence Detector (Beckman): determination of biophysical parameters (sedimentation coefficient, molar mass), dissociation constants (for Kd between pM and mM), stoichiometry and aggregation rate.
2) Analysis of biological interactions:
- PEAQ-ITC; ITC200 and VP-ITC (Malvern): measurements of protein-protein, protein-DNA, protein-ligand interactions to determine interaction-related parameters (stoichiometry, KD, deltaH).
- BLI (Octet Red96 - Sartorius) : BLI (Bio Layer Interferometry) is an optical technique for measuring macromolecular interactions by analysing the interference of white light reflected from the surface of a biosensor on which one of the partners is immobilised. BLI experiments are used to determine the kinetic parameters of the interaction (Kon and Koff) and the affinity (KD).
- HeliX (Dynamic Biosensors): switchSENSE® technology based on DNA nanolevials, electrically actuated to oscillate at high frequencies. Measurements of interactions between molecules (protein/DNA, protein/protein) making it possible to determine the kinetic parameters of the interaction (Kon and Koff) and to calculate the KD. Measurements of the size/conformation (DH) of a protein or macromolecular complex.
- MST Monolith NT.115 (NanoTemper): interaction measurements providing information on affinity constants (KD), with small amounts of sample, for a range of affinities between nM and mM. One of the partners must be labelled with a fluorophore (Alexa488 type).
3) Analysis of the stability of macromolecules and biological complexes, including formulation assistance:
- Auto PEAQ DSC and VP-DSC (Malvern): measurements of the stability of proteins, protein complexes, formulation aid. Technique for determining various thermodynamic parameters (Tonset, Tm, deltaHcal, deltaHvh).
- TSA (Applied Biosystem): determination of the thermal denaturation of proteins by measuring changes in SYPRO orange fluorescence.
- DSF : Tycho NT.6 (Nanotemper) : used in protein quality control: determination of thermal denaturation of proteins followed by intrinsic fluorescence variation (tryptophan and tyrosine).
Keywords
Offres de formations
Département(s) de recherche
- SDV (Life sciences)