Electrically induced transport properties for protein separation by chromatography

Subir Kumar Basak, Purdue University

Abstract

Electrical gradients enhance protein separations in chromatography columns. The goal of this dissertation was to improve a system in which application of an axial electric potential across a liquid chromatography column separates proteins without resorting to a chemical gradient. The research was based on the hypothesis that the size and charge of proteins in a dextran gel column can be moderated using an electric potential so that differences in individual retention, and therefore resolution would be enhanced. Application of a one-dimensional differential material balance that incorporated electrophoretic mobilities predicted protein retention as a function of the charge/mass ratio of proteins and the pore size of an electrically neutral stationary phase. An energy balance incorporating the thermal properties of dextran and Tris-glycine buffers enabled the calculation of radial temperature profiles. This theoretical analysis led to identification of conditions resulting in stable column operation at electrical potentials of 50 to 125 V/cm. Heat transfer and autothermal effects were controlled and a constant temperature maintained in a 15 mm i.d. column for over four hours. These mechanistic models enable the specification of flowrate, pH, buffer composition, mobile phase temperature, stationary phase porosity; and the polarity, duration, and field strength of the applied electric potential required to achieve protein separations in electrochromatography columns.

Degree

Ph.D.

Advisors

Ladisch, Purdue University.

Subject Area

Agricultural engineering|Analytical chemistry

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