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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...

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...

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Updated: Jul 5, 2026

A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples

A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples

Published on: October 1, 2009

Lectin affinity chromatography.

¹La Jolla Cancer Research Foundation, La Jolla, California, USA.

Current Protocols in Protein Science

|April 23, 2008

Summary

This summary is machine-generated.

This study details using lectins, such as Concanavalin A-Sepharose and Wheat Germ Agglutinin-agarose, for glycoprotein purification. The method involves binding glycoproteins to lectins via their sugar chains and eluting them with competing sugars.

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Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Published on: March 9, 2010

Area of Science:

Biochemistry
Protein Chemistry
Glycobiology

Background:

Glycoproteins are crucial in biological processes.
Efficient purification methods are essential for studying glycoproteins.
Lectins offer specific binding capabilities for carbohydrate moieties.

Purpose of the Study:

To describe a method for preparative glycoprotein purification using immobilized lectins.
To provide guidelines for small-scale testing and optimization of lectin-based purification.
To outline the fundamental principles of lectin affinity chromatography for glycoproteins.

Main Methods:

Utilizing immobilized lectins, specifically Concanavalin A-Sepharose and WGA-agarose.
Performing small-scale pilot procedures to assess lectin binding and elution.
Developing elution strategies using specific simple sugars that mimic lectin ligands.

Main Results:

Demonstrated the feasibility of using readily available lectin affinity matrices.
Established a protocol for determining optimal binding and elution conditions.
Confirmed the principle of selective glycoprotein capture and release based on carbohydrate interactions.

Conclusions:

Immobilized lectins provide a versatile and effective approach for glycoprotein purification.
The described method allows for tailored purification based on specific glycoprotein sugar structures.
This technique is adaptable for various scales of glycoprotein isolation.