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Related Concept Videos

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...
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...
Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
Silica particles offer advantages such as rigidity,...
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...
High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...

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Related Experiment Video

Updated: Jul 5, 2026

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization
10:41

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization

Published on: April 5, 2019

Ion-exchange chromatography.

A Williams1, V Frasca

  • 1Amersham Pharmacia Biotech, Piscataway, New Jersey, USA.

Current Protocols in Protein Science
|April 23, 2008
PubMed
Summary
This summary is machine-generated.

This guide details ion-exchange chromatography for protein separation, covering batch and column methods with gradient elution. It includes essential protocols for optimizing binding, elution, capacity, regeneration, and storage of ion-exchange media.

More Related Videos

Online Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline
11:09

Online Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline

Published on: January 5, 2017

Related Experiment Videos

Last Updated: Jul 5, 2026

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization
10:41

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization

Published on: April 5, 2019

Online Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline
11:09

Online Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline

Published on: January 5, 2017

Area of Science:

  • Biochemistry
  • Protein Chemistry
  • Chromatography

Background:

  • Ion-exchange chromatography is a fundamental technique for protein purification.
  • Effective separation requires careful optimization of binding and elution conditions.

Purpose of the Study:

  • To provide a comprehensive guide for planning and executing ion-exchange chromatography.
  • To detail methods for both batch and column chromatography with various elution gradients.
  • To offer support protocols for optimizing and maintaining the chromatography process.

Main Methods:

  • Describes batch adsorption and column chromatography techniques.
  • Explains the use of step and linear elution gradients (pH and salt concentration).
  • Includes protocols for pilot experiments, dynamic capacity calculation, gradient generation, media regeneration, and storage.

Main Results:

  • Provides a framework for determining optimal pH for binding and salt concentration for elution.
  • Enables calculation of the dynamic capacity of ion-exchange columns.
  • Offers methods for reproducible protein elution using continuous gradients.
  • Details procedures for regenerating and storing ion-exchange media for reuse.

Conclusions:

  • Successful protein separation via ion-exchange chromatography relies on meticulous planning and execution.
  • The outlined protocols facilitate efficient optimization, regeneration, and storage of chromatography media.
  • This unit serves as a practical resource for researchers performing protein purification using ion-exchange chromatography.