Preparative Electrochromatography of Proteins in Various Types of Porous Media

Published

March 1, 2000

Author(s)

C M. Tellez, Kenneth D. Cole

Abstract

The performance of commercial and enzymatically-modified size-exclusion (SE) gels in electrochromatography was compared for preparative protein separations. Dextran and agarose-based SE gels were subjected to enzymatic digestion under mild conditions. This treatment partially hydrolyzed the gel matrix modifying its pore-size distribution. Enzymatic treatment of agarose-based SE gels was found to increase the resolution of the separation. Successful separation of preparative amounts of the A and B forms of β-lactoglobulin (difference in electrophoretic mobility of 8.5 %) was achieved with a high degree of purity using agarose-based SE gels. The four major whey proteins, β-lactoglobulin, α-lactalbumin, BSA and immunoglobulins, were purified from an acid whey preparation. The degree of retention of a protein in electrochromatography followed their free-solution electrophoretic mobility ( ) when the protein was able to enter the gel pores and the ratio of diffusion/ when the protein was excluded.

Citation

Electrophoresis

Volume

21

Issue

No. 5

Pub Type

Journals

Keywords

agarase and dextranase digestion, preparative separation, protein electrochromatography, whey proteins, Β-lactoglobulin

Citation

Tellez, C. and Cole, K. (2000), Preparative Electrochromatography of Proteins in Various Types of Porous Media, Electrophoresis (Accessed May 13, 2024)