Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

The Equilibrium Binding Constant and Binding Strength

The Equilibrium Binding Constant and Binding Strength

The equilibrium binding constant (Kb) quantifies the strength of a protein-ligand interaction. Kb can be calculated as follows when the reaction is at equilibrium:

The Equilibrium Binding Constant and Binding Strength

The Equilibrium Binding Constant and Binding Strength

The equilibrium binding constant (Kb) quantifies the strength of a protein-ligand interaction. Kb can be calculated as follows when the reaction is at equilibrium:

Noncovalent Attractions in Biomolecules

Noncovalent Attractions in Biomolecules

Noncovalent attractions are associations within and between molecules that influence the shape and structural stability of complexes. These interactions differ from covalent bonding in that they do not involve sharing of electrons.
Four types of noncovalent interactions are hydrogen bonds, van der Waals forces, ionic bonds, and hydrophobic interactions.
Hydrogen bonding results from the electrostatic attraction of a hydrogen atom covalently bonded to a strong-electronegative atom like oxygen,...

Enzyme Kinetics

Enzyme Kinetics

Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Functionalization of a Rigid Divalent Ligand for LecA, a Bacterial Adhesion Lectin.

ChemistryOpen·2015

Same author

Thiourea-based spacers in potent divalent inhibitors of Pseudomonas aeruginosa virulence lectin LecA.

Organic & biomolecular chemistry·2015

Same author

Optimizing divalent inhibitors of Pseudomonas aeruginosa lectin LecA by using a rigid spacer.

Chemistry (Weinheim an der Bergstrasse, Germany)·2013

Same author

Tissue-type plasminogen activator binds to Aβ and AIAPP amyloid fibrils with multiple domains.

Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis·2013

Same author

Site-specific peptide and protein immobilization on surface plasmon resonance chips via strain-promoted cycloaddition.

Lab on a chip·2013

Same author

Unusual binding of Grb2 protein to a bivalent polyproline-ligand immobilized on a SPR sensor: intermolecular bivalent binding.

Biochimica et biophysica acta·2012

Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jun 15, 2026

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance (SPR)

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance (SPR)

Published on: November 29, 2014

Affinity constants for small molecules from SPR competition experiments.

Nico J de Mol¹

¹Department of Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Utrecht, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)

|March 11, 2010

Summary

This summary is machine-generated.

Directly measuring small molecules with surface plasmon resonance (SPR) is difficult. Competition assays provide an efficient alternative for determining binding constants of small molecules to analytes like antibodies.

More Related Videos

Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions

Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions

Published on: February 10, 2022

Rapid Determination of Antibody-Antigen Affinity by Mass Photometry

Rapid Determination of Antibody-Antigen Affinity by Mass Photometry

Published on: February 8, 2021

Related Experiment Videos

Last Updated: Jun 15, 2026

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance (SPR)

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance (SPR)

Published on: November 29, 2014

Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions

Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions

Published on: February 10, 2022

Rapid Determination of Antibody-Antigen Affinity by Mass Photometry

Rapid Determination of Antibody-Antigen Affinity by Mass Photometry

Published on: February 8, 2021

Area of Science:

Biochemistry
Analytical Chemistry
Biophysics

Background:

Directly assaying small molecules using surface plasmon resonance (SPR) is often challenging and requires laborious techniques.
Competition assays present a more practical and efficient approach for analyzing small molecule interactions.

Purpose of the Study:

To present a competition assay method for determining thermodynamic binding constants of small molecules interacting with analytes.
To demonstrate the adaptability of this method for analyzing interactions with bivalent analytes, such as antibodies.

Main Methods:

Immobilizing a known small ligand onto an SPR sensor surface.
Assaying the binding of a larger analyte in the presence of varying concentrations of the compounds under investigation.
Analyzing inhibition curves to derive thermodynamic binding constants for solution-phase interactions.

Main Results:

The competition assay method allows for the determination of binding constants for small molecules without individual immobilization.
The method is effective for analyzing interactions with both monovalent and bivalent analytes.
Observed differences in binding affinities between solution and SPR surface measurements are discussed.

Conclusions:

Competition assays offer a streamlined and versatile approach for characterizing small molecule-analyte interactions using SPR.
This method reduces the need for repetitive immobilization procedures, saving time and resources.
The findings contribute to a better understanding of molecular interactions in solution versus surface-bound states.