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Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

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...
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
Types Of Column Chromatography01:29

Types Of Column Chromatography

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...
Chromatography: Introduction01:10

Chromatography: Introduction

Chromatography is a technique used to separate compounds based on differences of partitioning between two phases, the stationary phase and the mobile phase.
The phase in which the compounds linger or on which the compounds adsorb is called the stationary phase, whereas the mobile phase is the solvent that carries the solutes to be analyzed. In traditional column chromatography, the mixture flows through the stationary phase, and the compounds partition between the stationary and mobile phases...
Ion Exchange01:17

Ion Exchange

Ion exchange chromatography separates charged molecules from a solution by reversibly exchanging them with mobile, or 'active', ions associated with the oppositely charged stationary phase. This method can be used to separate ions, soften and deionize water, and purify solutions. The polymers comprising the ion-exchange column are high-molecular-weight and chemically stable polymers, crosslinked to be porous and essentially insoluble. They are also functionalized with either acidic or basic...
Principles Of Column Chromatography01:13

Principles Of Column Chromatography

The chromatography technique was first invented in 1901 by Michael S. Tswett, a Russian botanist, to separate plant pigments using organic solvents. Further, in 1941, Archer John Porter Martin and R. L. M. Synge modified the technique by packing silica gel into a column. A mixture of amino acids was then separated on the packed column using chloroform and water mixture as the mobile phase. This was the first report on column chromatography. At present, column chromatography is a widely used...

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Updated: Jun 3, 2026

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

Published on: August 16, 2019

Some alternative coupling chemistries for affinity chromatography.

D S Pepper1

  • 1Scottish National Blood Transfusion Service, Edinburgh, UK.

Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2011
PubMed
Summary
This summary is machine-generated.

This study reviews methods for attaching ligands to solid phases for macromolecule purification. It highlights the need for critical evaluation of both published techniques and commercial products.

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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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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

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Last Updated: Jun 3, 2026

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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Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications
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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
06:25

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

Published on: April 26, 2016

Area of Science:

  • Biochemistry
  • Materials Science
  • Chemical Engineering

Background:

  • Numerous methods exist for immobilizing ligands onto solid supports.
  • These matrices are crucial for affinity purification of macromolecules.
  • Published literature and commercial product descriptions often emphasize strengths over weaknesses.

Purpose of the Study:

  • To critically assess various ligand-coupling methods for affinity purification matrices.
  • To provide a balanced perspective on the advantages and disadvantages of different approaches.
  • To guide researchers in selecting appropriate purification strategies.

Main Methods:

  • Literature review of published ligand-coupling techniques.
  • Analysis of commercial affinity matrix products.
  • Comparative evaluation of method efficacy and limitations.

Main Results:

  • A wide array of ligand immobilization strategies are available.
  • Enthusiastic reporting in publications and marketing can obscure practical limitations.
  • Understanding the nuances of each method is key for successful purification.

Conclusions:

  • Researchers should approach claims about purification methods with critical assessment.
  • A thorough understanding of coupling chemistry and matrix properties is essential.
  • Informed selection of affinity matrices enhances macromolecule purification efficiency.