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Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

1.4K
Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
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Types Of Column Chromatography01:29

Types Of Column Chromatography

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The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
When the...
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Silica Gel Column Chromatography: Overview01:10

Silica Gel Column Chromatography: Overview

1.8K
Silica gel column chromatography is a technique for separating compounds using a column packed with silica gel as the stationary phase. This method relies on differences in the polarity of compounds. Based on their polarities, compounds move between the stationary phase (silica gel) and the mobile phase (the solvent), forming discrete bands in the column.
Polar components tend to bind strongly to the silica gel, causing them to move slowly through the column. In contrast, nonpolar compounds...
1.8K
Ligand Binding Sites02:40

Ligand Binding Sites

13.6K
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...
13.6K
The Equilibrium Binding Constant and Binding Strength02:18

The Equilibrium Binding Constant and Binding Strength

14.0K
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:
14.0K

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Updated: Sep 22, 2025

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
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Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study

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Ligand Immobilization Methods for Affinity Chromatography.

Chandra K Dixit1

  • 1Lumos Diagnostics, Sarasota, FL, USA. chandrakumar.dixit@gmail.com.

Methods in Molecular Biology (Clifton, N.J.)
|May 18, 2022
PubMed
Summary
This summary is machine-generated.

Choosing the right chromatographic support is key for effective purification and quantification. Understanding resin chemistry and immobilization techniques ensures optimal ligand grafting for various applications.

Keywords:
Boronic acidConcanavalin ALigand immobilizationNickel-nitrilotriacetic acid (Ni-NTA)

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Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications
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Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
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Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications
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Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
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Area of Science:

  • Analytical Chemistry
  • Materials Science

Background:

  • A wide variety of chromatographic supports exist with diverse properties.
  • Proper selection of these materials is crucial for successful ligand immobilization.
  • Applications include purification, scavenging, and target quantification.

Purpose of the Study:

  • To explain common ligand immobilization techniques.
  • To guide the selection of appropriate chromatographic supports.
  • To provide a reference for commercially available resin functionalities.

Main Methods:

  • Explanation of ligand immobilization strategies.
  • Discussion of chromatographic support properties.
  • Compilation of data on commercially available resins.

Main Results:

  • Ligand immobilization techniques are detailed.
  • Key properties of chromatographic supports are highlighted.
  • Information on commercial resin functionalities is provided.

Conclusions:

  • Understanding resin chemistry is critical for optimizing ligand immobilization.
  • This chapter serves as a guide for selecting and utilizing chromatographic supports.
  • Effective immobilization enhances the performance of chromatographic applications.