Platform

Biophysics of Macromolecules and their Interactions

Patrick1

Head: Patrick England

Research

The Centre of Biophysics of Macromolecules and their Interactions (french acronym: PFBMI) federates an ensemble of state-of-the-art instrumentation for the in vitro molecular-scale characterization of biological macromolecules, together with well established technical and methodological expertise. The mission of the PFBMI is to provide an added value to a great variety of biological projects, enabling the characterization of the intrinsic properties of macromolecules and their assemblies (shape, folding, stability auto-association,…) and of the interactions in which they are involved (stoichiometry, thermodynamic and kinetic parameters, …).

Fields of expertise of the PFBMI :

Technologies available at the PFBMI :

The PFBMI also provides privileged access to instrumentation installed in other research units:

  • Differential scanning fluorimetry
  • Electrophoretic mobility
  • Infrared spectroscopy
  • Small-Angle X-ray Scattering (SAXS)

PFBMI equipments are available for the entire scientific community, national and international, but a priority is given to scientists from the Institut Pasteur in Paris and those from the Institut Pasteur International Network.
The PFBMI can welcome projects at any time through a single e-mail portal (biophysique@pasteur.fr). A project submission form (upload here) and a sample information form (upload here) must be attached.

tarifs

Services

Thermodynamical stability of macromolecules

Circular dichroism (CD) spectroscopy allows the determination of secondary and tertiary structures, the analyse of conformational changes, the kinetic of folding and the measure of thermal and chemical stability of folding. Differential Scanning Calorimeter (DSC) give access to the thermal stability of macromolecules and their complexes

Kinetic and thermodynamical properties of interactions

Static light scattering and viscometry  allow the measure of mass and intrinsic viscometry of macromolecules. It gives access to information on oligomerisation state and stœchiometry of assemblies. Analytical Ultracentrifugation made the measurement of mass and shape of macromolecules in solution. It gives access to oligomerisation state, stœchiometry and equilibrium constant of assemblies

Thermodynamical stability of macromolecules

Circular dichroism (CD) spectroscopy allows the determination of secondary and tertiary structures, the analyse of conformational changes, the kinetic of folding and the measure of thermal and chemical stability of folding. Differential Scanning Calorimeter (DSC) give access to the thermal stability of macromolecules and their complexes

Secondary and tertiary structures of macromolecules

Circular dichroism (CD), allows the measurement of secondary and tertiary structures, the analyse of conformational changes, the kinetic of folding and the thermal and chemical stability of folding Fluorescence spectroscopy  give access to the analysis of tertiary conformation, of thermal and chemical stability and of molecular interactions through the measure of fluorescence intensity, anisotropy and life-time. […]

Size and Shape of Macromolecules and their assemblies

Dynamic Light Scattering (cuvette or microplaque) allows to quantify the diffusion speed of macromolecules in solution and than to deduce their hydrodynamic radius.       Static Light Scattering and viscosimetry give access to mass and intrinsic viscosity of macromolecules, allowing to estimate the oligomerisation state and the stoechiometry of assemblies. Analytical Ultracentrifugation give access […]

Stoechiometry of macromolecular Assembly

Static Light Scattering allow to measure the mass and intrinsic viscosity of macromolecules Analytical Ultracentrifugation give access to the oligomerization state and the stoechiometry of the assemblies  

Equipments

Infrared spectroscopy (FTIR) • Jasco • 6100

Fourier Transform Infra Red (FTIR) spectroscopy allows to analyze the secondary structure of macromolecules by measuring absorption frequencies of some specific chemical groups (notably, for proteins that of the amide bond). Informations given by FTIR are complementary to those obtained bycircular dichroism and are more specifically suited for the precise study of the  ß content of proteins. The […]

Dynamic Light Scattering (DLS) • Wyatt • Dynapro Plate Reader

Studying the dynamic light scattering of particles in solution allows to quantify their diffusion coefficient and to deduce their hydrodynamic radius (which depends notably on their mass and shape). The PFBMI has a DynaPro Plate Reader (Wyatt), specially adapted for the study and the optimization of the homogeneity of samples of macromolecules in solution. This […]

Microscale Thermophoresis (MST) • Nanotemper • LabelFree

This technology notably allows to monitor a great variety of molecular interactions rapidly and with a very low sample consumption, by measuring optically the mobility of macromolecules in a microscopic temperature gradient generated in a capillary tube. The PFBMI has an instruments that detect intrinsic fluorescence of proteins (Monolith NT.LabelFree, Nanotemper).

Microcalorimetry Differential Scanning Calorimetry (DSC) • Malvern • CAP DSC

Microcalorimetry enables the thermodynamic characterization of molecular interactions in solution, and of biochemical reactions in general. Two types of microcalorimeters (ITC and DSC) are available at the PFBMI, depending on the parameters that one wishes to measure. Differential scanning calorimeter (DSC, Malvern) is principally dedicated to the study of the thermal stability of macromolecules and […]

Microcalorimetry • Malvern • ITC200

Microcalorimetry enables the thermodynamic characterization of molecular interactions in solution, and of biochemical reactions in general. Isothermal titration calorimeters (VP-ITC et ITC200), principally dedicated to the study of molecular interactions. By measuring the reaction heat at constant temperature, ITC allows the direct determination of binding stoichiometry (n), equilibrium constant (Ka), and variations of enthalpy (ΔH) […]

Analytical ultracentrifugation • Beckman-Coulter • ProteomeLab XL-I

The PFBMI has a ProteomeLab XL-I (Beckman-Coulter) ultracentrifuge with a dual UV/visible and interference detection system. The hydrodynamic behavior of a macromolecule and its sedimentation coefficient are determined by different parameters: volume, shape, density and mass. Analytical ultracentrifugation allows a direct measurement of the mass and sedimentation coefficient of macromolecules in solution. In the case […]

Dual polarization interferometry (DPI) • Farfield Biolin • Analight 4D

The Analight 4D (Farfield-Biolin) instrument, based on the Dual Polarization Interferometry (DPI) technology, allow to study simultaneously, in real time, the affinities and kinetic parameters of biomolecular interactions and the conformational or compaction changes that occur upon complex formation (going further than what is possible with surface plasmon resonance instruments). DPI is notably adapted to […]

Fluorescence Spectroscopy Stopped Flow • TgK Scientific • KinetAsyst SF61-DX2

Fluorescence spectroscopy is a versatile technique which allows to analyze, at steady-state or in real time, the variations of the environment of fluorescent probes, either intrinsic (for instance tryptophans in the case of proteins) or extrinsic (synthetic fluorophores). This technology can notably be applied to the study of the conformational stability of macromolecules, the characterization […]

Fluorescence Spectroscopy • Photon Technology International • Quantamaster QM4CW

Fluorescence spectroscopy is a versatile technique which allows to analyze, at steady-state or in real time, the variations of the environment of fluorescent probes, either intrinsic (for instance tryptophans in the case of proteins) or extrinsic (synthetic fluorophores). This technology can notably be applied to the study of the conformational stability of macromolecules, the characterization […]

Microscale Thermophoresis (MST) • Nanotemper • NT115

This technology notably allows to monitor a great variety of molecular interactions rapidly and with a very low sample consumption, by measuring optically the mobility of macromolecules in a microscopic temperature gradient generated in a capillary tube. The PFBMI has two instruments that detect  the fluorescence of extrinsinc synthetic fluorophores (Monolith NT.115 Blue/red , Nanotemper). […]

Surface Plasmon Resonance • Biorad • Proteon XPR36

The PFBMI has two microfluidic SPR instruments (Biacore T200, GE Healthcare, and ProteOn XPR36, BioRad) that allow the label-free analysis of molecular interactions in real time. To that avail, one of the interaction partners is immobilized, covalently or non-covalently, while the other partner is brought in contact with the sensorchip surface using a continuous buffer […]

Surface Plasmon Resonance • GE Healthcare • Biacore T200

The PFBMI has two microfluidic SPR instruments (Biacore T200, GE Healthcare, and ProteOn XPR36, BioRad) that allow the label-free analysis of molecular interactions in real time. To that avail, one of the interaction partners is immobilized, covalently or non-covalently, while the other partner is brought in contact with the sensorchip surface using a continuous buffer […]

Microcalorimetry • Malvern • VP ITC

Microcalorimetry enables the thermodynamic characterization of molecular interactions in solution, and of biochemical reactions in general. Isothermal titration calorimeters (VP-ITC et ITC200), principally dedicated to the study of molecular interactions. By measuring the reaction heat at constant temperature, ITC allows the direct determination of binding stoichiometry (n), equilibrium constant (Ka), and variations of enthalpy (ΔH) […]

Static Light Scattering and viscometry • Viscotek • TDA 302

The intensity of light scattering of a solution is directly proportional to the average molecular wieght and to the concentration of its components. The PFBMI has a multiple detector (Viscotek TDAmax, Malvern), coupled to a GPCmax size-exclusion chromatography system. Apart from a multi-angle light scattering detector, this instrument includes a refractometer/UV-visible spectrophotometer and a viscometer. […]

Dynamic Light Scattering • Wyatt • Dynapro MS800

Studying the dynamic light scattering of particles in solution allows to quantify their diffusion coefficient and to deduce their hydrodynamic radius (which depends notably on their mass and shape). The PFBMI has an instrument DynaPro MS800 (Wyatt), specially adapted for the study and the optimization of the homogeneity of samples of macromolecules in solution. This […]

Circular Dichroism • AVIV • 215

The PFBMI has an Aviv 215 instrument with accessories for the measure of total fluorescence and to perform titrations. Studying macromolecules (and in particular proteins) by circular dichroism allows to obtain information about their folding. For proteins, far UV (180-260 nm) and near UV (250-330 nm) circular dichroism measurements give insight respectively into their secondary […]

Publications

Contact

Phone: 01 45 68 86 09
Email: biophysique@pasteur.fr

Address
25-28 Rue du Docteur Roux
75015, Paris
France