Mechanistic description and experimental studies of electrochromatography of proteins

Abstract

Electrochromatography is a form of gradient liquid chromatography in which an

Preparative electrochromatography of proteins in various types of porous media

“…In the opposite case, when the electric field is applied such that electrophoretic mobility is in the same direction as convective flow, proteins would not be retarded. This is supported by our own results and those of Basak and Ladisch [12] who showed that when convection and electrophoresis are in the same direction, the elution time of b-Lg (a protein that penetrated gel pores) was not significantly affected by the presence of the field (i.e., there was no retention by concentration polarization).…”

“…They discussed thermal effects arising from Joule heating and described the separation of bovine hemoglobin and a-lactalbumin with an electric field of 80 V/cm and the partial separation of a-lactalbumin and b-lactoglobulin at 130 V/cm. High voltages (2000± 4000 V) and cooled columns have been used to separate other binary mixtures of proteins [12,13]. Cole and Cabezas [14] monitored the temperature increase in column eluent as a function of applied electric field and explored the effect of the gel degree of cross-linking in the EC of various proteins.…”

Retention behavior of proteins in size‐exclusion electrochromatography with a low‐voltage electric field perpendicular to the liquid phase streamline

“…During the past decades, many efforts have been made for the technical and theoretical development of preparative electrochromatography [1][2][3][4][5][6]. As a potential separation technique, electrokinetic transports, including electroosmosis and electrophoresis, are applied in electrochromatography to enhance mass transfer of solutes inside stationary phase particles and the liquid film around them, which are considered as the rate-limiting factors in the pressure-driven chromatographic processes [6,7].…”

“…As a potential separation technique, electrokinetic transports, including electroosmosis and electrophoresis, are applied in electrochromatography to enhance mass transfer of solutes inside stationary phase particles and the liquid film around them, which are considered as the rate-limiting factors in the pressure-driven chromatographic processes [6,7]. Currently, most preparative electrochromatography of proteins is generally established by applying an external electric field (eEF) in the longitudinal direction of the size-exclusion column [1][2][3][4][5]. In this case, eEF not only improves the mass transfer of electrolytes toward the solid matrices but also provides an additive driving force along the liquid phase streamline by electrokinetically driven transport.…”

Separation of different physical forms of plasmid DNA using a combination of low electric field strength and flow in porous media: Effect of different field gradients and porosity of the media

“…Solutes able to enter the porous media did not experience concentration polarization. Higher electric fields could be used to focus proteins on the column [11]. Analytical electrochromatography uses capillary columns, high electric fields, and solid (such as silica) chromatography materials to achieve rapid high-resolution separations [12±14].…”