Surface Plasmon Resonance Binding Assays

Stapled Peptide Affinity Evaluation and a-Helix Content Correlation 10.3.4.1 Surface Plasmon Resonance Binding Assays 

Surfece plasmon resonance (SPR) measures adsorption of material onto a planar metal (typically gold) surface and provides a very powerful tool for protein binding assays by analyzing the protein-ligand association and dissociation. Typically, pure protein is immobilized in buffer solution at different densities (according to the chips) using standard coupling reactions followed by ethanol-amine deactivation of the surfaces. A control surface is also prepared in the same manner in the absence of protein. 

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Uchida, FI., Fujitani, K., Kawai, Y., Kitazawa, H., Horii, A., Shiiba, K., Saito, K., and Saito, T. (2004). A new assay using surface plasmon resonance (SPR) to determine binding of the Lactobacillus acidophilus group to human colonic mucin. Biosci. Biotechnol. Biochem. 68, 1004 1010. 

The traditional methods of evaluating the interactions of glycans and glycan-binding proteins are based on isothermal titration calorimetry, surface plasmon resonance (SPR), and enzyme-linked lectin assays. Recently, glycan arrays have been developed as a... 

Zhou, V. et al. 2004. A time-resolved fluorescence resonance energy transfer-based HTS assay and a surface plasmon resonance-based binding assay for heat shock protein 90 inhibitors. Anal. Biochem. 331, 349-357. 

In the kinetic studies of the adsorption process, the mass transport of the analyte to the binding sites is an important parameter to account for. Several theoretical descriptions of the chromatographic process are proposed to overcome this difficulty. Many complementary experiments are now needed to ascertain the kinetic measurements. Similar problems are found in the applications of the surface plasmon resonance technology (SPR) for association rate constant measurements. In both techniques the adsorption studies are carried out in a flow system, on surfaces with immobilized ligands. The role of the external diffusion limitations in the analysis of SPR assays has often been mentioned, and the technique is yet considered as giving an estimate of the adsorption rate constant. It is thus important to correlate the SPR data with results obtained from independent experiments, such as those from chromatographic measurements. 

Wiltschi B, Knoll W, Sinner E-K (2006) Binding assays with artificial tethered membranes using surface plasmon resonance. Methods 39 134-146... 

Miao Y, Cui T, Leng F, Wilson WD (2008) Inhibition of high-mobility-group A2 protein binding to DNA by netropsin a biosensor-surface plasmon resonance assay. Anal Biochem 374 7-15... 

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