Quantitative monitoring of two simultaneously binding species using Label-Enhanced surface plasmon resonance
| Autor: | Brian V. Geisbrecht, Brandon L. Garcia, Lars H. Eng, Anders Hanning |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
education Biophysics Plasma protein binding Sensitivity and Specificity Biochemistry Article 03 medical and health sciences Protein structure Application areas Protein Interaction Mapping Molecule Binding site Surface plasmon resonance Coloring Agents Molecular Biology Binding Sites Staining and Labeling Chemistry Solid surface Proteins Reproducibility of Results Cell Biology Surface Plasmon Resonance Molecular Imaging Interaction studies Kinetics 030104 developmental biology Biological system Protein Binding |
| Zdroj: | Biochemical and Biophysical Research Communications. 497:133-138 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2018.02.040 |
| Popis: | Surface plasmon resonance (SPR) is a well-established method for biomolecular interaction studies. SPR monitors the binding of molecules to a solid surface, embodied as refractive index changes close to the surface. One limitation of conventional SPR is the universal nature of the detection that results in an inability to qualitatively discriminate between different binding species. Furthermore, it is impossible to directly discriminate two species simultaneously binding to different sites on a protein, which limits the utility of SPR, for example, in the study of allosteric binders or bi-specific molecules. It is also impossible in principle to discriminate protein conformation changes from actual binding events. Here we demonstrate how Label-Enhanced SPR can be utilized to discriminate and quantitatively monitor the simultaneous binding of two different species - one dye-labeled and one unlabeled - on a standard, single-wavelength SPR instrument. This new technique increases the versatility of SPR technology by opening up application areas where the usefulness of the approach has previously been limited. |
| Databáze: | OpenAIRE |
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