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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Advances in countercurrent chromatography for protein separations.

Nazim Mekaoui¹, Karine Faure, Alain Berthod

¹Laboratoire des Sciences nalytiques, Université de Lyon, ISA CNRS, Bat. CPE, 69622 Villeurbanne, France.

|April 20, 2012

Summary

Countercurrent chromatography (CCC) effectively purifies proteins using aqueous two-phase systems (ATPS). Hydrostatic CCC columns show significant potential for large-scale biological purification applications.

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Area of Science:

Biochemistry
Separation Science
Chromatography

Background:

Countercurrent chromatography (CCC) is a preparative purification method utilizing biphasic liquid systems.
Aqueous two-phase systems (ATPS) are increasingly recognized for their utility in biological purification processes.

Purpose of the Study:

This review focuses on protein separations achieved through CCC employing ATPS.
It aims to present the capabilities and equipment associated with this purification technique.

Main Methods:

The article discusses both hydrostatic and hydrodynamic CCC instruments.
It provides a comprehensive list of commercially available CCC equipment.
Key properties of ATPS are summarized, including phase diagrams and chemical compositions for polyethylene glycol/dipotassium phosphate/water systems.

Main Results:

Hydrostatic CCC columns demonstrate considerable potential for protein purification using ATPS aqueous phases.
Published protein purifications using this method have primarily occurred in academic settings.
A maximum throughput of 1.65 g/h of pure lysozyme was achieved using a 5.5 L hydrostatic CCC column.

Conclusions:

CCC, particularly with hydrostatic columns and ATPS, offers a promising approach for protein purification.
The technique has demonstrated scalability and efficiency, with potential for broader application in biological separations.