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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,...
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,...
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...

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Related Experiment Video

Updated: Jun 19, 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.

Alois Jungbauer1, Rainer Hahn

  • 1Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Vienna, Austria.

Methods in Enzymology
|November 7, 2009
PubMed
Summary

Ion-exchange chromatography is a popular protein separation technique. This method offers versatile purification under physiological conditions, enabling native protein recovery.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Protein Science

Background:

  • Ion-exchange chromatography (IEC) is a widely adopted technique for protein separation.
  • Its versatility makes it suitable for protein discovery, high-resolution purification, and industrial-scale protein production.
  • IEC operates under physiological salt and pH conditions, often preserving protein native structure.

Purpose of the Study:

  • To provide guidance on optimizing binding and elution conditions for ion-exchange chromatography.
  • To assist in the selection of appropriate stationary phases for effective protein separation.
  • To enhance the understanding and application of IEC in protein purification.

Main Methods:

  • The chapter focuses on the principles of ion-exchange chromatography.

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

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Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media
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Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media

Published on: May 13, 2022

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Last Updated: Jun 19, 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

Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media
10:46

Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media

Published on: May 13, 2022

  • Guidance is provided on manipulating salt gradients and pH for protein binding and elution.
  • Strategies for selecting suitable ion-exchange resins (stationary phases) are discussed.

    • Main Results:

    • Effective protein separation can be achieved using optimized IEC parameters.
    • The choice of stationary phase significantly impacts purification efficiency and resolution.
    • Native protein recovery is feasible under carefully controlled separation conditions.

    Conclusions:

    • Ion-exchange chromatography is a powerful and adaptable method for diverse protein purification needs.
    • Proper selection of binding/elution conditions and stationary phases is crucial for successful protein separation.
    • IEC facilitates both analytical and preparative-scale protein purification while maintaining protein integrity.