Separation of proteins from human plasma by sample displacement chromatography in hydrophobic interaction mode

• Published in: Electrophoresis Vol. 33; no. 12; pp. 1842 - 1849
• Main Authors: Josic, Djuro, Breen, Lucas, Clifton, James, Gajdosik, Martina Srajer, Gaso-Sokac, Dajana, Rucevic, Marijana, Müller, Egbert
• Format: Journal Article
• Language: English
• Published: Germany Blackwell Publishing Ltd 01.07.2012
• Subjects: Ammonium Sulfate; Blood Proteins - chemistry; Blood Proteins - isolation & purification; Chemical Precipitation; Chromatography, Liquid - methods; Databases, Protein; Human plasma; Humans; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Hydrophobic interaction; Polymers; Sample displacement chromatography; Sodium Chloride; Spectrometry, Mass, Electrospray Ionization; Therapeutic proteins
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.201200006

Sample displacement chromatography (SDC) in reversed‐phase and ion‐exchange modes was introduced approximately 20 years ago. This method was first used for the preparative purification of peptides and proteins. Recently, SDC in ion‐exchange mode was also successfully used for enrichment of low‐abundance proteins from human plasma. In this paper, the use of SDC for the separation of plasma proteins in hydrophobic interaction mode is demonstrated. By use of two or more columns coupled in series during sample application, and subsequent elution of detached columns in parallel, additional separation of bound proteins was achieved. Further low‐abundance, physiologically active proteins could be highly enriched and detected by ESI‐MS/MS.