Equipment

Biacore 3000

SPR analysis, which depends on surface immobilization of one of the interacting molecules, is performed on a Biacore 3000 instrument. In addition to affinity constants (KD), this technique also allows determination of kinetic rate constants ka and kd (association and dissociation rates).

Applications

Surface Plasmon Resonance, on the Biacore platform, is most frequently used for binding/kinetic analysis of interactions between protein-protein or protein-small molecule. However, it can detect binding between any kinds of molecules, provided that one of them (termed the ligand in SPR terminology) can be immobilized on the chip surface either covalently or non-covalently. As listed below, SPR has also been employed for a number of other applications.

Binding analysis - binding or no binding

Kinetic analysis - determination of affinity (KD) and rate constants ka and kd

• Concentration measurements - determination of analyte concentration

• Identification of interaction partners - bound analyte protein can be recovered and identified by mass spectrometry

• Epitope mapping

Highlights

• Low sample consumption. Often a few micrograms of both analyte and ligand is sufficient for a full kinetics experiment.

• Kinetic parameters ka and kd can be obtained.

• A broad range of affinities (mM to pM).

• Ligand needs to be immobilized, either covalent or non-covalently.