Version :
Integrated Structural Biology Grenoble
Accueil > SPR (Biacore technology) > SPR (Biacore technology)

Contact person(s) related to this article / CHOUQUET Anne / REISER Jean-Baptiste

SPR (Biacore technology)

Presentation

The Biacore technology allows real-time detection and monitoring of biomolecular binding events. In a Biacore experiment, one of the interacting molecules (the ligand) is bound to the biosensor surface (sensor chip), whereas the other (the analyte) is delivered to the surface in a continuous buffer flow through a microfluidic system. Binding to the immobilized molecule is followed by surface plasmon resonance (SPR), which detects mass changes at the sensor surface. Recording SPR signal variation as a function of time (sensorgram) for several analyte concentrations allows to determine the association and dissociation rate constants, and to derive the affinity constant. This technology also allows to measure the concentration of functional molecules and the interaction stoichiometry.
The Biacore technology is used to characterize molecular interactions involving small molecules, proteins, polysaccharides, lipids and nucleic acids. Sequential injections of several interactants make possible the mapping of interaction domains and the determination of the composition and of the mechanism of assembly of multiprotein complexes.

The IBS Biacore facility provides 2 instruments.

The following sections describe the functioning and use of this facility.

Dedicted staff

The scientific and technical expertises, and the management of the facility are provided by :

Responsibility:
Jean-Baptiste Reiser

Management:
Anne Chouquet

Equipement

  • 1 Biacore T200 : automatic injections, 4 flowcells simultaneously
  • 1 Biacore 3000 : automatic injections, 4 flowcells simultaneously

The Biacores T200 and 3000 are under maintenance contract with 1 to 2 annual preventive visits by a service engineer.
The computers are under local maintenance (infogérance CEA).

Access

  • open to academics according to term to define (service or collaboration).
  • open to industrials on a service basis and depending on the availability of the instruments.

Contact and localisation

The platform is located in the IBS building, room 515

Jean-Baptiste Reiser

Anne Chouquet

Use of the facility

Each novel user must contact Jean-Baptiste Reiser. A preliminary meeting is needed to discuss of the experimental conditions and to schedule the use of the machine in order to define the better strategy to carry out the study.

Each novel user will get trained and agrees to comply with the general conditions of use and to follow the operating instructions.

Each user can access the Biacore availability via the following calendars :

Trained users, provided that they have been trained and have accepted the general conditions of use, have to send a message to booking-spr psb-grenoble.eu stating the booking period and the Biacore of their choice. Their reservation will then be recorded.

The platform staff is available to users for advice and practical assistance during use of the machines.

Data processing

Each user is responsible for analyzing his data using the BiaEval software available on each instrument. Data transfer in text format is possible for later analysis using other softwares.
The users of each apparatus Biacore 3000 and Biacore T200 have to save the data acquired during their experiences either by the mean of removable external memory devices (USB drive, external hard drive, recommended for non-IBS users) or by the mean of the access to the IBS internal network and data storage system (recommended for IBS users). The instrument control computers are connected to IBS internal network to allow printing and data storage. The platform does not guarantee any data storage, archiving and recovery in case of loss following an informatics problem.

Cost

Each user provides his sensor chips, buffers and his consumables specific of his analysis.
For the pricing of the SPR platform please contact us.
Anne Chouquet
Jean-Baptiste Reiser

Follow up / Development

The platform staff helps users as requested in the technical choices (buffers, sensorchips) and in data processing and analysis.

Users must mention the platform in their publications: “We thank Isabelle Bally, Nicole Thielens and Jean-Baptiste Reiser. This work used the platforms of the Grenoble Instruct centre (ISBG; UMS 3518 CNRS-CEA-UJF-EMBL) with support from FRISBI (ANR-10-INSB-05-02) and GRAL (ANR-10-LABX-49-01) within the Grenoble Partnership for Structural Biology (PSB)."
The collaborative projects imply that the person responsible for the platform is a co-author of the published papers.

The SPR platform is engaged in developing quality management certification steps wich led in June 2011 for certification according to ISO 9001:2008

Web site

GE Healthcare Europe GmbH

Parc Technologique

Rue René Razel

Saclay

91898 Orsay Cedex, France.

Publications / References

General information on the 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 (data obtained using the instruments of the IBS platform)

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.