Surface Plasmon Resonance (SPR)

Highly sensitive analysis of binding phenomena in real-time without the need for labeling or size limitations

Nicole Thielens Nicole Thielens Institut de Biologie Structurale email hidden

Instruct has 4 centres offering Surface Plasmon Resonance (SPR) across Europe. Navigate the map and click on the pins to discover centres near you.

Instruct Centre - CERM/CIRMMP Italy
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Instruct Centre - France 2
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Protein facility - NKI
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Surface Plasmon Resonance (SPR) Details

In cellular structural biology the analysis of biomolecular interactions is crucial for describing interaction networks and monitoring multiple complex formation. Surface Plasmon Resonance (SPR) allows a highly sensitive analysis of binding phenomena in real-time without the need for labeling or size limitations. SPR can be used for determining equilibrium constants (Kd), stoichiometry, and association (ka) and dissociation (kd) rates in weak and strong interacting systems, and has applications in molecular assembly, epitope mapping, and small molecule screening. In SPR experiments, one of the interacting moieties is immobilized onto the biosensor surface and the other is free in the solution that is made to flow over the biosensor surface. The response in SPR signal is proportional to the mass of the material bound to the biosensor surface, so large interacting molecules give a more intense signal than small ones. The availability of specific sensor chips for different application purposes makes SPR a powerful tool in structural biology and a basic resource for Instruct.

User Guide

SPR is a technique for measuring the binding between interactants (e.g. proteins) in real time. Usually one protein is covalently linked on a sensor chip, while a second protein (or any other molecule) is made to flow across the surface of the chip.

SPR can be used to investigate several biomolecular interactions:

  • Protein-protein
  • Protein-nucleic acid
  • Protein-lipid
  • Protein-carbohydrate
  • Protein-small molecule

SPR analysis can provide answers to several questions:

  • How strong is the interaction (KD)?
  • How fast is the interaction (ka or kon)?
  • How fast is complex dissociation (kd or koff)?

SPR analysis can be performed at a wide range of buffers, pH, and temperatures.

A few micrograms of ligand (the immobilized partner) per chip surface are required. Samples and buffers must be filtered through a 0.2 micron filter, and de-gassed before use.

The measurement consists of the following steps:

  • Chip activation
  • Ligand immobilization
  • Analyte injection
  • Data analysis

All of the components required for interaction analysis are available at the facility:

  • Instrumentation
  • Software for analysis
  • Sensor chips
  • Buffers
  • Reagent kits and protocol development kits