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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...

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...

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Related Experiment Video

Updated: Dec 3, 2025

Preparation of Highly Porous Coordination Polymer Coatings on Macroporous Polymer Monoliths for Enhanced Enrichment of Phosphopeptides

Preparation of Highly Porous Coordination Polymer Coatings on Macroporous Polymer Monoliths for Enhanced Enrichment of Phosphopeptides

Published on: July 14, 2015

Macroporous Polymer Monoliths for Affinity Chromatography and Solid-Phase Enzyme Processing.

E G Korzhikova-Vlakh¹, G A Platonova¹, T B Tennikova²

¹Institute of Macromolecular Compounds, Russian Academy of Sciences, St. Petersburg, Russia.

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

|October 31, 2020

Summary

Macroporous monoliths enable fast separation and purification of biomolecules using affinity chromatography. These versatile materials also support enzyme immobilization for various biotechnological applications.

Keywords:

Affinity chromatography Conjoint chromatography Enzyme reactors Enzymes Fractionation Monoliths Protein and peptide immobilization Proteins

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Last Updated: Dec 3, 2025

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Low Molecular Weight Protein Enrichment on Mesoporous Silica Thin Films for Biomarker Discovery

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Published on: April 17, 2012

Area of Science:

Biotechnology
Analytical Chemistry
Biochemistry

Background:

Monolithic stationary phases offer high permeability and unique morphology for efficient mass transfer processes.
High-performance liquid chromatography (HPLC) on monoliths, particularly in bioaffinity mode, is ideal for rapid separation of biomacromolecules and supramolecular systems.

Purpose of the Study:

To present applications of macroporous monoliths in modern affinity processing.
To detail methods for isolating specific biomolecules and immobilizing enzymes.

Main Methods:

Utilizing commercially available macroporous monoliths for affinity separation and purification.
Developing multifunctional monolithic columns for complex antibody fractionation.
Immobilizing various enzymes onto macroporous monolith supports.

Main Results:

Demonstrated efficient isolation of monospecific antibodies from rabbit serum.
Achieved simultaneous affinity separation of Protein G and serum albumin from human serum.
Successfully isolated recombinant products like Protein G and tissue plasminogen activator.
Showcased multifunctional antibody fractionation using stacked monolithic columns.
Adapted monoliths for immobilizing diverse enzymes, creating functional immobilized enzyme reactors.

Conclusions:

Macroporous monoliths are highly effective for fast and efficient biomolecule separation and purification via affinity chromatography.
Monoliths provide a versatile platform for enzyme immobilization, enabling the development of advanced enzyme reactors for biotechnological applications.