A novel stationary phase derivatized from hydrophilic gigaporous polystyrene-based microspheres for high-speed protein chromatography

J Chromatogr A. 2009 Sep 11;1216(37):6511-6. doi: 10.1016/j.chroma.2009.07.059. Epub 2009 Aug 3.

Abstract

Using agarose coated gigaporous polystyrene microspheres as a base support, a novel anion exchanger (DEAE-AP) has been developed after functionalization with diethylaminoethyl chloride. The gigaporous structure, static adsorption behavior, and chromatographic properties of DEAE-AP medium were characterized and compared with those of commercially available resin DEAE Sepharose Fast Flow (DEAE-FF). The results implied that there existed some through pores in DEAE-AP microspheres, which effectively reduced resistance to stagnant mobile phase mass transfer by inducing convective flow of mobile phase in the gigapores of medium. As a consequence, the column packed with DEAE-AP exhibited low column backpressure, high column efficiency, high dynamic binding capacity and high protein resolution at high flow velocity up to 2600cm/h. In conclusion, all the results suggested that the gigaporous absorbent is promising for high-speed protein chromatography.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid / instrumentation*
  • Chromatography, Ion Exchange
  • Ethanolamines
  • Microspheres*
  • Polystyrenes*
  • Proteins / analysis
  • Proteins / isolation & purification*
  • Sepharose / analogs & derivatives
  • Sepharose / ultrastructure
  • Technology, Pharmaceutical

Substances

  • Ethanolamines
  • Polystyrenes
  • Proteins
  • Sepharose
  • 2-diethylaminoethanol