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.

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 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.

Mechanism of the regulation of type IB phosphoinositide 3OH-kinase byG-protein βγ subunits

2002 ◽  
Vol 362 (3) ◽  
pp. 725-731 ◽  
Author(s):  
Sonja KRUGMANN ◽  
Matthew A. COOPER ◽  
Dudley H. WILLIAMS ◽  
Phillip T. HAWKINS ◽  
Len R. STEPHENS
Keyword(s):  
Plasma Membrane ◽  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Lipid Vesicles ◽  
Dissociation Rate ◽  
Lipid Monolayers ◽  
Dissociation Rate Constants

Type IB phosphoinositide 3OH-kinase (PI3K) is activated by G-protein βγ subunits (Gβγs). The enzyme is soluble and largely cytosolic in vivo. Its substrate, PtdIns(4,5)P2, and the Gβγs are localized at the plasma membrane. We have addressed the mechanism by which Gβγs regulate the PI3K using an in vitro approach. We used sedimentation assays and surface plasmon resonance to determine association of type IB PI3K with lipid monolayers and vesicles of varying compositions, some of which had Gβγs incorporated. Association and dissociation rate constants were determined. Our results indicated that in an assay situation in vitro the majority of PI3K will be associated with lipid vesicles, irrespective of the presence or absence of Gβγs. In line with this, a constitutively active membrane-targeted PI3K construct could still be activated substantially by Gβγs in vitro. We conclude that Gβγs activate type IB PI3K by a mechanism other than translocation to the plasma membrane.

Compound Selectivity and Target Residence Time of Kinase Inhibitors Studied with Surface Plasmon Resonance

2017 ◽  
Vol 429 (4) ◽  
pp. 574-586 ◽  
Author(s):  
Nicole Willemsen-Seegers ◽  
Joost C.M. Uitdehaag ◽  
Martine B.W. Prinsen ◽  
Judith R.F. de Vetter ◽  
Jos de Man ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Residence Time ◽  
Plasmon Resonance ◽  
Kinase Inhibitors

Compound selectivity and target residence time of kinase inhibitors studied with surface plasmon resonance

2016 ◽  
Vol 69 ◽  
pp. S79
Author(s):  
N. Willemsen-Seegers ◽  
J. Uitdehaag ◽  
M. Prinsen ◽  
J. De Vetter ◽  
J. De Man ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Residence Time ◽  
Plasmon Resonance ◽  
Kinase Inhibitors

Characterization of the Carbohydrate Binding Specificity and Kinetic Parameters of Lectins by Using Surface Plasmon Resonance

1999 ◽  
Vol 274 (2) ◽  
pp. 203-210 ◽  
Author(s):  
Simon R. Haseley ◽  
Philippe Talaga ◽  
Johannis P. Kamerling ◽  
Johannes F.G. Vliegenthart
Keyword(s):  
Surface Plasmon Resonance ◽  
Kinetic Parameters ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Binding Specificity ◽  
Carbohydrate Binding

scholarly journals Kinetic Analysis and Epitope Mapping of Monoclonal Antibodies to Salmonella Typhimurium Flagellin Using a Surface Plasmon Resonance Biosensor

Antibodies ◽  
2019 ◽  
Vol 8 (1) ◽  
pp. 22 ◽  
Author(s):  
Devendra Bhandari ◽  
Fur-Chi Chen ◽  
Shreya Hamal ◽  
Roger Bridgman
Keyword(s):  
Monoclonal Antibodies ◽  
Surface Plasmon Resonance ◽  
Salmonella Typhimurium ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Epitope Mapping ◽  
Dissociation Constants ◽  
Interaction Model ◽  
Dissociation Rate ◽  
Surface Plasmon Resonance Biosensor

Salmonella Typhimurium is one of the leading causes of foodborne diseases worldwide. Biosensors and immunoassays utilizing monoclonal antibodies are widely used for the detection and subtyping of S. Typhimurium. However, due to insufficient information on the nature of binding with S. Typhimurium flagellin, the selection of appropriate antibodies for assay development is a cumbersome task. Hence, we aimed to compare the binding kinetics of a panel of monoclonal antibodies and their relative binding sites to flagellin antigen using a surface plasmon resonance biosensor. Initially, the flagellin was captured on the sensor surface through an immobilized anti-flagellin antibody. The interactions of different concentrations of monoclonal antibodies to flagellin were determined, and binding curves were fitted using 1:1 bio-interaction model to calculate the kinetic parameters. For epitope mapping, pairwise comparisons were completed to determine the binding inhibition of each paired combination of monoclonal antibodies. It was found that these monoclonal antibodies differed significantly (p < 0.05) in association rate, dissociation rate, and equilibrium dissociation constants. Of the five monoclonal antibodies, only two interfered with the binding of each other. Four distinct epitopes located within a 23 kDa domain of flagellin were identified. Findings from this study provide crucial information needed for the further development and optimization of biosensors and other immunoassays for the detection and subtyping of Salmonella.

Surface plasmon resonance study for a reliable determination of the affinity constant of multivalent grafted beads

Soft Matter ◽  
2021 ◽  
Author(s):  
Giuseppina Simone
Keyword(s):  
Surface Plasmon Resonance ◽  
Kinetic Parameters ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Biological Samples ◽  
Affinity Constant ◽  
Cluster Effect ◽  
Reliable Determination ◽  
Thermodynamic And Kinetic Parameters

Thermodynamic and kinetic parameters of the multivalent galactose beads are determined via SPR. The results show that the glycoside cluster effect influences the affinity toward the lectin and it is crucial for mimicking the biological samples.

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