Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Affinity Chromatography

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 Chromatography

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...

Extraction: Advanced Methods

Extraction: Advanced Methods

Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is formed in...

Ion Exchange

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...

Types Of Column Chromatography

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...

Matrix-Assisted Laser Desorption Ionization (MALDI)

Matrix-Assisted Laser Desorption Ionization (MALDI)

Matrix-assisted laser desorption ionization (MALDI) is a powerful analytical technique used in mass spectrometry. It enables the identification and characterization of various biomolecules, including proteins, peptides, nucleic acids, and carbohydrates. MALDI is an ionization technique, widely employed in biological and medical research, as well as in fields like pharmacology and biochemistry.The analyte of interest, a biomolecule or a mixture of biomolecules, is mixed with a suitable matrix...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Erratum to: 10th asilomat conference on mass spectrometry: Time of flight mass spectrometry.

Journal of the American Society for Mass Spectrometry·2013

Same author

Thiophilic adsorption chromatography.

Methods in molecular biology (Clifton, N.J.)·2011

Same author

Serum amyloid A is elevated in the serum of lung cancer patients with poor prognosis.

British journal of cancer·2010

Same author

Remarkable application of serum EBV EBER-1 in monitoring response of nasopharyngeal cancer patients to salvage chemotherapy.

Annals of the New York Academy of Sciences·2001

Same author

Direct evidence of the generation in human stomach of an antimicrobial peptide domain (lactoferricin) from ingested lactoferrin.

Biochimica et biophysica acta·1999

Same author

Prognostic significance of DNA flow cytometric analysis in patients with nasopharyngeal carcinoma.

Cancer·1998

Search research articles

Related Experiment Video

Updated: Jun 3, 2026

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography

Published on: January 17, 2020

Immobilized metal ion affinity chromatography.

T T Yip¹, T W Hutchens

¹Department of Pediatrics, Baylor College of Medicine, Houston, TX.

Methods in Molecular Biology (Clifton, N.J.)

|March 25, 2011

Summary

Immobilized metal ion affinity chromatography (IMAC) offers a versatile separation technique by chelating metal ions to stationary phases for selective protein binding. This method bridges specific bioaffinity and broader adsorption techniques, enabling diverse applications in protein purification.

More Related Videos

Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory

Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory

Published on: August 23, 2018

Ion Mobility-Mass Spectrometry Techniques for Determining the Structure and Mechanisms of Metal Ion Recognition and Redox Activity of Metal Binding Oligopeptides

Ion Mobility-Mass Spectrometry Techniques for Determining the Structure and Mechanisms of Metal Ion Recognition and Redox Activity of Metal Binding Oligopeptides

Published on: September 7, 2019

Related Experiment Videos

Last Updated: Jun 3, 2026

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography

Published on: January 17, 2020

Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory

Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory

Published on: August 23, 2018

Ion Mobility-Mass Spectrometry Techniques for Determining the Structure and Mechanisms of Metal Ion Recognition and Redox Activity of Metal Binding Oligopeptides

Ion Mobility-Mass Spectrometry Techniques for Determining the Structure and Mechanisms of Metal Ion Recognition and Redox Activity of Metal Binding Oligopeptides

Published on: September 7, 2019

Area of Science:

Biochemistry
Analytical Chemistry
Separation Science

Background:

Immobilized metal ion affinity chromatography (IMAC) is a chromatography technique also known as metal chelate chromatography.
It functions as an intermediate separation method between highly specific bioaffinity separations and less specific adsorption methods like ion exchange.
IMAC stationary phases are engineered to chelate metal ions, which then selectively bind to specific amino acid residues (e.g., histidine) on proteins and peptides.

Purpose of the Study:

To provide an overview of Immobilized Metal Ion Affinity Chromatography (IMAC) as a versatile protein separation technique.
To highlight the design principles and capabilities of IMAC stationary phases.
To showcase the scope and adaptability of IMAC through examples and a list of synthesis procedures.

Main Methods:

Development of stationary phases designed for chelating specific metal ions.
Utilizing the selective binding affinity of chelated metal ions for amino acid residues (e.g., His) on target proteins.
Exploration of various immobilized chelating groups, including iminodiacetate (IDA), tris(carboxymethyl) ethylenediamine (TED), and metal-binding peptides.

Main Results:

IMAC stationary phases can be synthesized with unlimited potential for efficient metal ion chelation.
The critical factor for IMAC effectiveness is sufficient exposure of the metal ion for biospecific interaction with proteins.
A comprehensive list of published procedures for synthesizing and utilizing IMAC stationary phases is presented, demonstrating its versatility.

Conclusions:

Immobilized metal ion affinity chromatography is a powerful and adaptable technique for protein and peptide separation.
The continuous exploration of novel immobilized chelating groups enhances the capabilities of IMAC.
IMAC serves as a valuable tool bridging highly specific and broadly selective separation methods in biochemistry and biotechnology.