scholarly journals Surface plasmon resonance: a useful technique for cell biologists to characterize biomolecular interactions

2013 ◽  
Vol 24 (7) ◽  
pp. 883-886 ◽  
Author(s):  
Robert V. Stahelin
Keyword(s):  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Protein Interactions ◽  
Plasmon Resonance ◽  
Dissociation Rate ◽  
Biomolecular Interactions ◽  
Protein Protein Interactions ◽  
Interaction Kinetics ◽  
Lipid Protein Interactions ◽  
Dissociation Rate Constants

Surface plasmon resonance (SPR) is a powerful technique for monitoring the affinity and selectivity of biomolecular interactions. SPR allows for analysis of association and dissociation rate constants and modeling of biomolecular interaction kinetics, as well as for equilibrium binding analysis and ligand specificity studies. SPR has received much use and improved precision in classifying protein–protein interactions, as well as in studying small-molecule ligand binding to receptors; however, lipid–protein interactions have been underserved in this regard. With the field of lipids perhaps the next frontier in cellular research, SPR is a highly advantageous technique for cell biologists, as newly identified proteins that associate with cellular membranes can be screened rapidly and robustly for lipid specificity and membrane affinity. This technical perspective discusses the conditions needed to achieve success with lipid–protein interactions and highlights the unique lipid–protein interaction mechanisms that have been elucidated using SPR. It is intended to provide the reader a framework for quantitative and confident conclusions from SPR analysis of lipid–protein interactions.

scholarly journals Quick Evaluation of Kinase Inhibitors by Surface Plasmon Resonance Using Single-Site Specifically Biotinylated Kinases

2013 ◽  
Vol 19 (3) ◽  
pp. 453-461 ◽  
Author(s):  
Daisuke Kitagawa ◽  
Masaki Gouda ◽  
Yasuyuki Kirii
Keyword(s):  
Surface Plasmon Resonance ◽  
Kinetic Parameters ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Kinase Inhibitors ◽  
Growth Factor Receptor ◽  
Biochemical Properties ◽  
Dissociation Rate ◽  
Single Site ◽  
Dissociation Rate Constants

In evaluating kinase inhibitors, kinetic parameters such as association/dissociation rate constants are valuable information, as are equilibrium parameters KD and IC50 values. Surface plasmon resonance (SPR) is a powerful technique to investigate these parameters. However, results are often complicated because of impaired conformations by inappropriate conditions required for protein immobilization and/or heterogeneity of the orientation of immobilization. In addition, conventional SPR experiments are generally time-consuming. Here we introduce the use of single-site specifically biotinylated kinases combined with a multichannel SPR device to improve such problems. Kinetic parameters of four compounds—staurosporine, dasatinib, sunitinib, and lapatinib—against six kinases were determined by the ProteOn XPR36 system. The very slow off-rate of lapatinib from the epidermal growth factor receptor and dasatinib from Bruton’s tyrosine kinase and colony stimulating factor 1 receptor (CSF1R) were confirmed. Furthermore, IC50 values were determined by an activity-based assay. Evaluating both physicochemical and biochemical properties would help to understand the detailed character of the compound.

Identification of protein-protein interactions using Protein Microarrays and Surface Plasmon Resonance Measurements

2005 ◽  
Vol 2005 (Fall) ◽  
Author(s):  
Harald Seitz ◽  
Claus Hultschig ◽  
Holger Eickhoff ◽  
Carsten Zeilinger ◽  
Ulrike Borgmeier ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Protein Interactions ◽  
Plasmon Resonance ◽  
Protein Microarrays ◽  
Protein Protein Interactions

scholarly journals Surface plasmon resonance unveils important pitfalls of enzyme-linked immunoassay for the detection of anti-infliximab antibodies in patients’ sera

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Marten Beeg ◽  
Cesare Burti ◽  
Eleonora Allocati ◽  
Clorinda Ciafardini ◽  
Rita Banzi ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Dissociation Rate ◽  
Therapeutic Antibodies ◽  
Serum Concentrations ◽  
High Affinity ◽  
Bowel Diseases ◽  
Inflammatory Bowel ◽  
Dissociation Rate Constants

AbstractMeasurements of serum concentrations of therapeutic antibodies and anti-drug antibodies (ADA) can support clinical decisions for the management of non-responders, optimizing the therapy. In the present study we compared the results obtained by classical ELISA and a recently proposed surface plasmon resonance (SPR)-based immunoassay, in 76 patients receiving infliximab for inflammatory bowel diseases. The two methods indicated very similar serum concentrations of the drug, but there were striking differences as regards ADA. All the sera showing ADA by ELISA (14) also showed ADA by SPR, but the absolute amounts were different, being 7–490 times higher with SPR, with no correlation. Eight patients showed ADA only with SPR, and these ADA had significantly faster dissociation rate constants than those detectable by both SPR and ELISA. The underestimation, or the lack of detection, of ADA by ELISA is likely to reflect the long incubation steps which favor dissociation of the patient’s low-affinity ADA, while the commercial, high-affinity anti-infliximab antibodies used for the calibration curve do not dissociate. This problem is less important with SPR, which monitors binding in real time. The possibility offered by SPR to detect ADA in patients otherwise considered ADA-negative by ELISA could have important implications for clinicians.

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