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Integrated Structural Biology Grenoble
Accueil > SPR

Contacts relatifs à cet article / CHOUQUET Anne / REISER Jean-Baptiste

Surface Plasmon Resonance

PRESENTATION

The SPR/BLI platform provides a BIAcore/GE Healthcare life sciences instrument for the characterization of biomolecular interactions by Surface Plasmonic Resonance (SPR) detection.
SPR is an optical technique that measures changes in the refractive index on the surface of a metal surface. As they are dependent on the nature and mass of the constituents on the surface, this technique allows the detection in real time and without labelling of biological molecules.

With the Biacore T200, the technology is applied to the study of interactions between partners and the determination of kinetic constants of associations and dissociations (kon, koff), affinity constant (KD) or thermodynamic constants (dH, tdS).
The method is based on a biosensor on which a first molecule (the ligand) is immobilized, and the binding of the second partner (the analyte) is measured during its injection by a microfluidic system in a continuous flow of buffer onto the biosensor surface. The response is measured in real time (sensorgram) and is directly proportional to the mass density of the fixed biomolecules (1000 RU <-> 1 ng/mm2 <-> 10 mg/ml).
It applies to the characterization of molecular interactions involving small molecules and all classes of biological macromolecules (proteins, polysaccharides, lipids and nucleic acids).

ACCESS, LOCATION, BOOKING, RATES AND CONTACTS

Access
The SPR platform is open to :

  • Academics according to modalities to be defined (access to the service/autonomous or collaboration) according to the involvement of the platform staff.
  • Industrial in the form of a service on quotation.

Tariffs
The access tariffs are available for consultation here.
The rates are exclusive of tax (excluding VAT) and per day of use. They can be reviewed once a year.
The prices include access to one of the 2 instruments of the platform and to the equipment and consumables described in the paragraph "The platform provides". They do not include specific consumables that will be provided by the user.
Any additional consumables provided by the platform and used will be charged in addition to the price of access to the instrument.
On request, a quotation can be provided.

Location
The SPR platform is located on the EPN campus in Grenoble, in the IBS building, room 515 : http://www.isbg.fr/contact/article/access
External users must request authorization to access the EPN campus. Contact the platform staff before your visit.
External users can request accommodation at EPN campus Guest house (at their own expense). Contact the platform staff before your visit.

Use of the platform
New users
Each new user must contact the platform staff before any experimentation or instrument reservation.
A preliminary meeting is mandatory for :

  • Define the terms of use of the platform : collaboration or autonomous access.
  • Discuss the experimental conditions and define the best strategy for carrying out the study.
  • Schedule training and first use of the instrument.

Each user must agree to comply with the GENERAL CONDITIONS AND OPERATING INSTRUCTIONS.

Referred users
Trained and registered users can book directly after checking the booking schedule.

Availability and reservation
Booking
Each user must reserve their use of the Biacore T200 by filling in the RESERVATION FORM.

Availability
The Availability of the Biacore T200 is available here


Contacts
Address
SPR/BLI platform
Institute of Structural Biology - UMR 5075 (CNRS-CEA-UJF)
71 Avenue des Martyrs
CS 10090
38044 Grenoble cedex 9
FRANCE
E-mail : ibs-plateforme-spr.contact ibs.fr

Scientific Manager
Jean-Baptiste Reiser
E-mail : jean-baptiste.reiser ibs.fr
Telephone : +33 (0)4 57 42 42 42 85 49

Technical and administrative assistance
Anne Chouquet
E-mail : anne.chouquet ibs.fr
Telephone : +33 (0)4 57 42 85 85 85 47

GETTING STARTED GUIDE

A usage guide for the Biacore T200 is downloadable : GETTING STARTED GUIDE.

WHAT TO BRING FOR YOUR EXPERIMENTS

You need

  • Running buffer(s) in 1 L glass bottle.
  • Ligand samples at the highest concentrations (to be diluted in immobilization buffers).
  • Analytes samples at the highest concentrations (to be diluted in running buffers).
  • Regeneration buffers if known.
  • Biosensors/Sensochips series-S
  • Pipettes and tips (preferably checked and calibrated).
  • Necessary plastic tubes (Falcon, Eppendorf type tubes).
  • Any previous and relevant data and/or protocols.

Ordering consumables and kits
Users must provide consumables specific to their experiments.
Consumables must be ordered from Biacore/GE Healthcare Life Sciences (www.biacore.com).
No other biosensors/sensochips will be accepted.
For the Biacore T200, biosensor/sensorchips must be Series-S type.

WHAT THE PLATFORM PROVIDES

The SPR platform provides basic and common supplies for experiments on the Biacore T200 :

  • Biacore T200 instrument.
  • 4°C refrigerator and -20°C -20°C -20°C freezer for sample storage.
  • Vortex and benchtop centrifuge.
  • P20/Tween-20 (added to your experiment buffers)
  • Immobilization kits for amine coupling methods.
  • Regeneration scouting kit.
  • Tubes and caps for Biacore T200.
  • External users who cannot bring laboratory supplies can borrow supplies from our laboratory (pipettes, tips, plastic and glass items).
  • If necessary, a UV/visible nanodrop spectrophotometer is available in our laboratory (room 520).
  • Filtered water solutions, instrument maintenance.

HOW MUCH TIME DO I NEED ?

The time required to fully characterize your interaction depends on your biological system, the strategy used, the time needed to optimize your experiences and the difficulties you may encounter...... It can vary from a few days to a few weeks.....
Here are some estimates of the time required for traditional methods :

  • Starting sequence : 20 to 30 min.
  • pH Scouting before immobilization by amine coupling : 30 to 60 min per ligand sample.
  • Immobilization by amine coupling : 45 to 60 min per sample.
  • Secondary capture : 20 to 30 min per sample.
  • Pilot analyses : ½ daily to several days depending on the number of sample injections at different concentrations, testing of several contact and dissociation times, several flow rates and regeneration conditions.
  • MCK/Steady State : 3 to 8 hours or more depending on kinetics and concentrations.
  • SCK : 2 to 5 hours depending on the kinetics.
  • Stop sequence : 30 min.

COMPUTING

The platform provides computers dedicated to the control of each instrument, the implementation of experiments, data acquisition and recording and data analysis,
The platform provides an additional computer with free access for data analysis and processing,
All computers are equipped with regularly updated software necessary for data monitoring and analysis,

ACKNOWLEDGEMENTS, PUBLICATIONS AND VALORIZATION

Collaborations
Collaborative projects require that platform staff co-author publications in peer-reviewed journals and other communications in which SPR or BLI data appear and in accordance with the terms previously discussed.

Service - made available
Users undertake to mention the platform and its staff in all publications or communications in which SPR or BLI data appear by mentioning the following sentence : "This work used the platforms of the Grenoble Instruct-ERIC center (ISBG ; UMS 3518 CNRS-CEA-UGA-EMBL) within the Grenoble Partnership for Structural Biology (PSB), supported by FRISBI (ANR-10-INBS-05-02) and GRAL, financed within the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003). Authors acknowledge the SPR/BLI platform personal, Jean-Baptiste REISER Ph.D. and Anne Chouquet, for their help and assistance."

Quality
As part of its structuring and continuous improvement, the SPR technical platform is committed to the implementation of a quality approach that led to ISO 9001:2015 certification since June 2011.

REFERENCES

Generalities on Biacore technology

Homola J. Surface plasmon resonance sensors for detection of chemical and biological species. Chem. Rev. 2008 ; 108:462-93.

Karlsson R., Katsamba P.S., Nordin H., Pol E., Myszka D.G. Analyzing a kinetic titration series using affinity biosensors. Anal. Biochem. 2006 ; 349:136-47.

Karlsson R., Larsson A. Affinity measurement using surface plasmon resonance. Methods Mol. Biol. 2004 ; 248:389-415.

Karlsson R. SPR for molecular interaction analysis : a review of emerging application areas. J. Mol. Recognit. 2004 ; 17:151-61.

Publications
(works realised on platform instruments)

Bally I, Ancelet S, Moriscot C, Gonnet F., Mantovani A, Daniel R, Schoen G, Arlaud GJ.,Thielens NM, (2013) Expression of recombinant human complement C1q allows identification of the C1r/C1s-binding sites. PNAS 110, 8650-8655

Rossi V., Bally I., Ancelet S., Xu Y., Frémeaux-Bacchi V., Vivès R.R., Sadir R., Thielens N., Arlaud G.J. (2010) Functional characterization of the recombinant human C1 inhibitor serpin domain : insights into heparin binding. J. Immunol. 184, 4982-4989.

Gout E., Garlatti V., Smith D.F., Lacroix M.M., Dumestre-Perard C., Lunardi T., Martin L., Cesbron J.-Y., Arlaud G.J., Gaboriaud C., Thielens N.M. (2010) Carbohydrate recognition properties of human ficolins : Glycan array screening reveals the sialic acid binding specificity of M-ficolin. J. Biol. Chem. 285, 6612-6622.

Lacroix M., Dumestre-Pérard C., Schoehn G., Houen G., Cesbron J.-Y., Arlaud G.J., Thielens N.M. (2009) Residue Lys57 in the collagenous region of human L-ficolin and its counterpart Lys47 in H-ficolin play a key role in the interaction with the MASPs and the collectin receptor calreticulin. J. Immunol. 182, 456-465.

Baleux F., Loureiro-Morais L., Hersant Y., Clayette P., Arenzana-Seisdedos F., Bonnaffé B. Lortat-Jacob H.(2009) A synthetic CD4-HS glycoconjugate inhibits both CCR5 and CXCR4 HIV-1 attachment and entry. Nat. Chem. Biol. 5, 743-748.

Gras S., Saulquin X., Reiser J.-B., Debeaupuis E., Echasserieau K., Kissenpfennig A., Legoux F., Chouquet A., Le Gorrec M., Machillot P., Neveu B., Thielens N., Malissen B., Bonneville M., Housset D. (2009) Structural bases for the affinity driven selection of a public TCR against a dominant human cytomegalovirus epitope. J. Immunol. 183, 430-437.

Crublet E., Andrieu J.-P., Vivès R.R., Lortat-Jacob H. (2008) The HIV-1 envelope glycoprotein gp120 features four heparan sulfate binding domains, including the coreceptor binding site. J. Biol. Chem. 283, 15193-15200.

Attali C., Frolet C., Durmort C., Offant J., Vernet T., Di Guilmi A.M : (2008) Streptococcus pneumoniae choline-binding protein E interaction with plasminogen/plasmin stimulates migration across the extracellular matrix. Infect. Immun. 76, 466-476.

Sattin S., Daghetti A., Thepaut M., Berzi A., Sánchez-Navarro M., Tabarani G., Rojo J., Fieschi F., Clerici M., Bernardi A.(2010) Inhibition of DC-SIGN-mediated HIV infection by a linear trimannoside mimic in polyvalent presentation ACS Chemical Biology 5, 301-312.

Timpano G., Tabarani G., Anderluh M., Invernizzi D., Vasile F., Potenza D., Nieto P., Rojo J., Fieschi F., Bernardi A. (2008) Synthesis of novel DC-SIGN ligands with an ?-fucosylamide anchor ChemBioChem, 9, 1921-30.