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Related Concept Videos

Protein-Drug Binding: Determination Methods01:22

Protein-Drug Binding: Determination Methods

Determining protein-drug binding can be achieved through indirect and direct methods, each providing valuable insights into the interaction between proteins and drugs.
Indirect methods involve isolating the bound drug from its free form in biological samples such as blood, serum, or plasma. These techniques aim to measure the percentage of drugs bound to proteins. Equilibrium dialysis is a commonly used method where the free drug concentration at equilibrium is measured by separating the bound...
The Equilibrium Binding Constant and Binding Strength02:18

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:
Protein-protein Interfaces02:04

Protein-protein Interfaces

Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a polypeptide...
Conserved Binding Sites01:49

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...
Conserved Binding Sites01:49

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...
Ligand Binding Sites02:40

Ligand Binding Sites

Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...

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Related Experiment Video

Updated: Jun 20, 2026

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
15:27

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

Published on: April 17, 2017

Updates on contemporary protein binding techniques.

Victor Tuan Giam Chuang1, Toru Maruyama, Masaki Otagiri

  • 1School of Pharmacy, Curtin University of Technology, Perth, Australia.

Drug Metabolism and Pharmacokinetics
|September 12, 2009
PubMed
Summary

High-throughput techniques are crucial for protein binding analysis. A novel assay using lipid membranes shows promise for accurately assessing drug-protein binding, even for challenging lipophilic compounds.

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Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

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Protein Purification-free Method of Binding Affinity Determination by Microscale Thermophoresis
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Protein Purification-free Method of Binding Affinity Determination by Microscale Thermophoresis

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Last Updated: Jun 20, 2026

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
15:27

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

Published on: April 17, 2017

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions
06:01

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

Published on: January 7, 2019

Protein Purification-free Method of Binding Affinity Determination by Microscale Thermophoresis
10:22

Protein Purification-free Method of Binding Affinity Determination by Microscale Thermophoresis

Published on: August 15, 2013

Area of Science:

  • Biochemistry
  • Pharmacology
  • Analytical Chemistry

Background:

  • Protein binding analysis is vital for drug discovery and development.
  • Existing methods for determining protein binding and affinity vary in complexity and application.
  • High-throughput screening is essential for efficient drug development pipelines.

Purpose of the Study:

  • To review current techniques for protein binding analysis.
  • To introduce a novel high-throughput assay for protein binding determination.
  • To evaluate the assay's suitability for challenging drug compounds.

Main Methods:

  • Review of established protein binding assays (e.g., ultrafiltration, affinity chromatography).
  • Discussion of advanced techniques like surface plasmon resonance and photoaffinity labeling.
  • Introduction of a new high-throughput assay utilizing solid-supported lipid membranes (Transil).

Main Results:

  • Established methods often require separation or measure changes in protein properties.
  • Newer techniques offer improved specificity and site identification.
  • The Transil assay provides valid results, including for strongly bound and lipophilic drugs.

Conclusions:

  • Automated high-throughput techniques are indispensable for advancing protein binding analysis.
  • The novel Transil assay is effective for determining protein binding, particularly for lipophilic drugs.
  • This method has the potential to accelerate drug discovery and development through efficient protein binding determination.