Surface Plasmon Resonance (SPR)

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

 
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Surface Plasmon Resonance (SPR) Details

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

Technical Specifications

Different immobilization strategies can be used:

  • Amine coupling via Lys and Arg residues of proteins
  • Biotin-streptavidin coupling
  • Thiol coupling
  • Maleimide coupling
  • NTA-NTA functionalized chip for capturing HIS-tagged proteins

SPR has a wide range of applicability:

  • MW >100 Da (analyte)
  • kd 103-107 M-1s-1
  • ka10-5- 0.5 s-1
  • KD~ 10-3 -10-12 M