Ion-Exchange Sample Displacement Chromatography as a Method for Fast and Simple Isolation of Low- and High-Abundance Proteins from Complex Biological Mixtures

Martina Srajer Gajdosik1, Spomenka Kovac1, Nela Malatesti2, Egbert Müller3
and Djuro Josic2,4*

Department of Chemistry, Josip Juraj Strossmayer University, Cara Hadrijana 8/A,
HR-31000 Osijek, Croatia

2Department of Biotechnology, University of Rijeka, Radmile Matejčić 2, HR-51000 Rijeka, Croatia
3Tosoh Bioscience GmbH, Zettachring 6, DE-70567 Stuttgart, Germany
4Warren Alpert Medical School, Brown University, Richmond Street 222, RI-02903 Providence, USA

Article history:

Received April 18, 2013

Accepted December 18, 2013

Key words:

sample displacement chromatography, ion-exchange mode, plasma proteins


Sample displacement chromatography (SDC) in reversed phase and ion-exchange
modes was introduced at the end of 1980s. This chromatographic method was first used for preparative purification of synthetic peptides, and subsequently adapted for protein fractionation, mainly in anion-exchange mode. In the past few years, SDC has been successfully used for enrichment of low- and medium-abundance proteins from complex biological fluids on both monolithic and bulk chromatographic supports. If aqueous mobile phase is used with the application of mild chromatographic conditions, isolated proteins are not denatured and can also keep their biological activity. In this paper, the use of SDC in anion-exchange mode on a high-capacity chromatographic resin for separation of proteins from complex biological mixtures such as human plasma is demonstrated. By use of three and more columns coupled in series during sample application, and subsequent parallel elution of detached columns, additional separation of bound proteins was achieved. Highly enriched human serum albumin fraction and a number of physiologically active medium- and low-abundance proteins could be fractionated and detected by electrospray ionization tandem mass spectrometry (ESI-MS/MS) and matrix-assisted laser desorption/ ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF-MS). The use of the aforementioned columns that can be sanitized with 1 M sodium hydroxide for further application of SDC in biotechnology and food technology was discussed.


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