Oscillatory transverse electric field enhances mass transfer and protein capacity in ion-exchange electrochromatography.
Ion-exchange electrochromatography with an oscillatory electric field perpendicular to mobile-phase flow driven by pressure (pIEEC) was developed with a column design of rectangle cross-section. The effect of electric field strength on the dynamic binding capacity (DBC) was examined by frontal analysis of bovine serum albumin (BSA) adsorption to the packed beds of DEAE Sepharose FF in Tris-glycine buffer (pH 8.2). It was shown that the DBC at 10% breakthrough (Q(10)) in the pIEEC increased linearly with increasing the electric field strength. For example, with a packed-bed height of 15mm and electric potential gradient of 38V/cm, Q(10) increased four times over that in normal ion-exchange chromatography. So, the transverse electric field has created significant electro-kinetic mass transports (electroosmosis and electrophoresis) that intensified exterior liquid-film and intraparticle mass transfers, leading to the increased protein binding capacity. Due to the increased capacity in the pIEEC, partial resolution of BSA and IgG under an overload condition was realized without any process optimization. The results have revealed that an electric potential gradient of 20V/cm was enough to greatly enhance the DBC in the pIEEC, and when necessary, high electric field strength can be realized with a low applied voltage because the side distance of the column is usually an order of magnitude smaller than its height. The use of low voltage to carry out electrochromatography is a significant advantage of the pIEEC over conventional electrochromatography with axial electric field.
Tan
Guo-Min
GM
Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
Shi
Qing-Hong
QH
Sun
Yan
Y
D000327:Adsorption; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004553:Electric Conductivity; D004586:Electrophoresis / Q000379:methods*; D007136:Immunoglobulins / Q000302:isolation & purification; D007700:Kinetics; D009995:Osmosis; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification
D007136:Immunoglobulins; D011506:Proteins; D012710:Serum Albumin, Bovine