Protein adsorption in two-dimensional electrochromatography packed with superporous and microporous cellulose beads
Dongmei WANG, Guodong JIA, Liang XU, Xiaoyan DONG, Yan SUN
Protein adsorption in two-dimensional electrochromatography packed with superporous and microporous cellulose beads
Anion-exchange superporous cellulose (DEAE-SC) and microporous cellulose (DEAE-MC) adsorbents were packed in an electrochromatographic column, and the effect of external electric field (eEF) on the dynamic adsorption was investigated. The column was designed to provide longitudinal, transverse or 2-dimensional (2D) eEF. It was found that the electro-kinetic effect caused by the introduction of an electric field played an important role in the dynamic adsorption of bovine serum albumin to the adsorbents. The dynamic binding capacity (DBC) in the presence of 2D eEF was higher than in the presence of a one-dimensional eEF. The effect of flow velocity on the DBC of the two adsorbents was also demonstrated. It was found that the effect of electric field on the DEAE-MC column was more remarkable than that on the DEAE-SC column at the same flow rate, whereas the DEAE-SC column showed higher DBC and adsorption efficiency (AE) than the DEAE-MC column. With increasing flow rate, the DEAE-SC column could still offer high DBC and AE in the presence of the 2D eEF. For example, a DBC of 21.4 mg/mL and an AE of 57.7% were obtained even at a flow rate as high as 900 cm/h. The results indicate that the 2D electrochromatography packed with the superporous cellulose adsorbent is promising for high-speed protein chromatography.
electrochromatography / two-dimensional electric field / dynamic binding capacity / superporous cellulose bead / protein
[1] |
Afeyan N B, Reginer F E, Dean R C. Perfusive chromatography. <patent>US Patent, 5019270</patent>, 1991
|
[2] |
Harrison R G, Todd P W, Rudge S R, Petrides D. Bioseparation Science and Engineering. New York: Oxford University Press, 2003
|
[3] |
Afeyan N B, Fulton S P. Perfusion chromatography packing materials for proteins and peptides. J Chromatogr, 1991, 544: 267–279
CrossRef
Google scholar
|
[4] |
Gustavsson P E, Axelsson A, Larsson P O. Superporous agarose beads as a hydrophobic interaction chromatography support. J Chromatogr A, 1999, 830: 275–284
CrossRef
Google scholar
|
[5] |
Shi Q H, Zhou X, Sun Y. A novel superorous agarose medium for high-speed protein chromatography. Biotechnol Bioeng, 2005, 92: 643–651
CrossRef
Google scholar
|
[6] |
Wang D M, Hao G, Shi Q H, Sun Y. Fabrication and characterization of superporous cellulose bead for high-speed protein chromatography. J Chromatogr A, 2007, 1146: 32–40
CrossRef
Google scholar
|
[7] |
Liapis A I, Grimes B A. Film Mass transfer coefficient expressions for electroosmotic flows. J Colloid Interface Sci, 2000, 229: 540–543
CrossRef
Google scholar
|
[8] |
Rudge S R, Basal S K, Ladisch M R. Solute retention in electrochromatography by electrically induced sorption. AIChE J, 1993, 39: 797–808
CrossRef
Google scholar
|
[9] |
Keim C, Ladisch M. New system for preparative electrochromatography of protins. Biotechnol Bioeng, 2002, 70: 72–81
CrossRef
Google scholar
|
[10] |
Liu Z, Yin G, Feng S H, Wang D H, Ding F X, Yuan N J. Oscillatory electroosmosis-enhanced intra/inter-particle liquid transport and its primary applications in the preparative electrochromatography of proteins. J Chromatogr A, 2001, 921: 93–98
CrossRef
Google scholar
|
[11] |
Tan G M, Shi Q H, Sun Y. Retention behavior of proteins in size-exclusion electrochromatography with a low-voltage electric field perpendicular to the liquid phase streamline. Electrophoresis, 2005, 26: 3084–3093
CrossRef
Google scholar
|
[12] |
Tan G M, Shi Q H, Sun Y. Oscillatory transverse electric field enhances mass transfer and protein capacity in ion-exchange electrochromatography. J Chromatogr A, 2005, 1098: 131–137
CrossRef
Google scholar
|
[13] |
Jia G D, Sun Y. Dye-ligand affinity electrochromatography with transverse and/or longitudinal electric fields. Sep Purif Technol, 2008, 59: 277–285
CrossRef
Google scholar
|
[14] |
Shi Y, Sun Y. Fabrication and characterization of a novel biporous spherical adsorbent for protein chromatography. Chromatographia, 2003, 57: 29–35
CrossRef
Google scholar
|
[15] |
Sun G Y, Yang Z, Dong X Y, Sun Y. Bioporous polymeric beads fabricated by double emulsification for high-speed protein chromatography. J Appl Polym Sci, 2007, 103: 17–23
CrossRef
Google scholar
|
[16] |
Yarmush M L, Olson W C. Electrophoretic elution from biospecific adsorbents: principles, methodology, and applications. Electrophoresis, 1988, 9: 111–120
CrossRef
Google scholar
|
[17] |
Horstmann B J, Chase H A. Modeling the affinity adsorption of immunoglobulin G to protein A immobilized to agarose matrices. Chem Eng Des, 1989, 67: 243–254
|
[18] |
Skidmore G L, Horstmann B J, Chase H A. Modeling single-component protein adsorption to the cation exchanger Sepharose FF. J Chromatogr A, 1990, 498: 113–128
CrossRef
Google scholar
|
[19] |
Rathore A S, Horvath C. Capillary electrochromatography: theories on electroosmotic flow in porous media. J Chromatogr A, 1997, 781: 185–195
CrossRef
Google scholar
|
[20] |
Grimes B A, Meyers J J, Liapis A I. Determination of the intraparticle electroosmotic volumetric flow-rate, velocity and Peclet number in capillary electrochromatography from pore network theory. J Chromatogr A, 2000, 890: 61–72
CrossRef
Google scholar
|
[21] |
Leinweber F C, Pfafferodt M P, Seidel-Morgenstern A, Tallarek U. Electrokenetic effects on the transport of charged analytes in biporous media with discrete ion-permselective regions. Anal Chem, 2005, 77: 5839–5850
CrossRef
Google scholar
|
/
〈 | 〉 |