A novel stationary phase derivatized from hydrophilic gigaporous polystyrene-based microspheres for high-speed protein chromatography.
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.
Qu
Jian-Bo
JB
National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, China.
Wan
Xing-Zhong
XZ
Zhai
Yan-Qin
YQ
Zhou
Wei-Qing
WQ
Su
Zhi-Guo
ZG
Ma
Guang-Hui
GH
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D002852:Chromatography, Ion Exchange; D004983:Ethanolamines; D008863:Microspheres; D011137:Polystyrenes; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification*; D012685:Sepharose / Q000031:analogs & derivatives / Q000648:ultrastructure; D013678:Technology, Pharmaceutical
D004983:Ethanolamines; D011137:Polystyrenes; D011506:Proteins; D012685:Sepharose; C007369:2-diethylaminoethanol