title: Preparative electrochromatography of proteins in various types of porous media.
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 beta-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, beta-lactoglobulin, alpha-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 (mu) when the protein was able to enter the gel pores and the ratio of diffusion/mu when the protein was excluded.
authors.affiliation: Mechanical and Industrial Engineering Department, University of Illinois at Urbana-Champaign, Urbana, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Tellez
authors.forename: K D
authors.initials: KD
authors.lastname: Cole
mesh_terms: D002845:Chromatography / Q000379:methods*; D004058:Diffusion; D004586:Electrophoresis / Q000379:methods*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006868:Hydrolysis; D007136:Immunoglobulins / Q000302:isolation & purification; D007768:Lactalbumin / Q000302:isolation & purification; D007782:Lactoglobulins / Q000302:isolation & purification; D008894:Milk Proteins / Q000302:isolation & purification; D012685:Sepharose / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000302:isolation & purification; D000067816:Whey Proteins
chemical_list: D007136:Immunoglobulins; D007782:Lactoglobulins; D008894:Milk Proteins; D000067816:Whey Proteins; D012710:Serum Albumin, Bovine; D012685:Sepharose; D007768:Lactalbumin; D006026:Glycoside Hydrolases; C019624:agarase