FRISBI

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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

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SOUTH PARIS

SPRI

The team of « Dynamics of Macromolecular Complexes» of the LBPA located at the ENS Cachan has been developing and using label free techniques to study macromolecular interactions involving nucleoproteins, antigen-antibody, peptide-peptide and peptide-proteins for many years. The main approach has involved Surface Plasmon Resonance (SPR) and Surface Plasmon Resonance Imagery (SPRi). The group now offers a platform that provides customised chips to study almost any type of macromolecular interaction.

SPR consists of immobilizing ligands to a surface and then observing changes in the refractive index at the surface as molecules bind. In the SPRi technique initially introduced by Genoptics (now Horiba) the biosensor surface consist of prisms made of a high refractive index material with one surface coated with a thin layer of gold. An evanescent field called a plasmon wave is created at the interface of this gold-coated surface and the dielectric from a light beam arriving through the prism at an angle of total internal reflection. At this angle there is a resonance effect that is measured by imaging the entire reflected light from a monochromatic polarized electroluminescent diode using a CCD camera linked via a dedicated optical system. This allows analysis of an entire surface upon which discrete spots of ligand have been immobilized.

Real time analysis provides information about the on (ka) and off (kd) kinetic rates for a macromolecular interaction between a molecule in solution and an immobilised ligand (and thus the Kd) and stoichiometry. A major difference between the SPR (as exemplified by BIAcore) and SPRi apart from the obvious advantage of multiplexing in the latter is that SPRi allows the development of specific monolayer surfaces.  LBPA has developed a series of specific surface chemistries (General Liquid Interface Specific Surfaces –GLISS) that allow macromolecules to be favourably immobilised and orientated in a Self Assembled Monolayer that is extremely refractive to non-speciifc binding. Up to 400 independently immobilised different molecules may be interogated simultaneously on the same surface. For immobilisation each spot uses less than 2 µl of material at concentrations ranging from 0.01 to 0.1 mg/ml, and a range of amide/carboxyl/thiol coupling chemistries are available.  Volumes of between 10 and 500 µl may be flowed across the surfaces at flow rates of between 10 and 500 µl/min.

 

The platform consists of :

Bibliography

1)     Emeline Bouffartigues, Hervé Leh, Marielle Anger-Leroy, Sylvie Rimsky and Malcolm Buckle, Rapid coupling of Surface Plasmon Resonance (SPR and SPRi) and ProteinChip™ based mass spectrometry for the identification of proteins in nucleoprotein interactions. (2007) Nucleic Acids Research. 2007; 35(6):e39. Epub 2007 Feb 7.

2)     Malcolm Buckle. Plasmon Resonance revisited. Bioworld Europe (2008) 01.  Pp18-21

3)     Prieto IKouznetsova AFütterer ATrachana VLeonardo EAlonso Guerrero ACano Gamero MPacios-Bras C, Leh H, Buckle M, Garcia-Gallo M, Kremer LSerrano ARoncal FAlbar JPBarbero JLMartínez-A Cvan Wely KH. Synaptonemal complex assembly and histone trimethylation determine DIDO3 localization in meiosis Chromosoma. (2009) Oct;118(5):617-32. Epub 2009 Jun 26

4)     Soufi ANoy PBuckle MSawasdichai AGaston KJayaraman PS. Sheela CK2 phosphorylation of the PRH/Hex homeodomain functions as a reversible switch for DNA binding.  Nucleic Acids Res. (2009) Jun;37(10):3288-300

5)     Claude Nogues, Hervé Leh, Christopher G. Langendorf, Ruby H.P. Law, Ashley M. Buckle and Malcolm Buckle. Characterisation of Peptide Microarrays for Studying Antibody-Antigen Binding Using Surface Plasmon Resonance Imagery PLoS One. 2010 Aug 13;5(8):e12152. doi: 10.1371/journal.pone.0012152

6)     Claude Nogues, Hervé Leh, Joseph Lautru, Olivier Delelis and Malcolm Buckle Efficient antifouling surface for quantitative Surface Plasmon Resonance based biosensor analysis. PLoS One. 2012;7(9):e44287. doi: 10.1371/journal.pone.0044287. Epub 2012 Sep 12.

7)     Paulina Prorok, Christine Saint Pierre, Didier Gasparuttoc, Olga S. Federova, Alexander A. Ischenko, Hervé Leh, Malcolm Buckle, Barbara Tudek and Murat Saparbaev. Highly mutagenic exocyclic DNA adducts are substrates for the human nucleotide incision repair pathway. PLoS One. 2012;7(12):e51776. doi: 10.1371/journal.pone.0051776. Epub 2012 Dec 14

8)     Elvire Beleoken, Herve Leh, Claude Nogues, Eleonora De Martin, Catherine Johanet, Didier Samuel, Jean-Charles Duclos-Vallée, Malcolm Buckle, Eric Ballot. SPRi-based strategy to identify specific biomarkers in systemic lupus erythematosus, rhumatoid arthritis and autoimmune hepatitis. PloS 1 2013; 8 (12) e84600