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Oscillatory transverse electric field enhances protein resolution and capacity of size‐exclusion chromatography
Author(s) -
Tan GuoMin,
Dong XiaoYan,
Sun Yan
Publication year - 2006
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200500354
Subject(s) - chemistry , chromatography , electric field , ovalbumin , myoglobin , electrochromatography , analytical chemistry (journal) , size exclusion chromatography , lysozyme , protein purification , resolution (logic) , capillary electrochromatography , stationary phase , biochemistry , physics , enzyme , immune system , quantum mechanics , artificial intelligence , computer science , immunology , biology
Protein separations by a novel size‐exclusion electrochromatography (SEEC) are presented. The present SEEC, denoted as pSEEC, was established with an oscillatory low‐voltage electric field perpendicular to the mobile‐phase streamline. Retention experiments with different proteins indicated that the influence of electric field strength on the partition coefficient is different for different proteins as well as for the same protein under different mobile‐phase conditions. These results of protein retention led to the experimental design of protein separations with binary mixtures of BSA and immunoglobulin G (IgG), myoglobin (Myo) and lysozyme (Lys), as well as ovalbumin (Oval) and Myo. The separation results for the binary protein systems sufficiently exhibited the applicability of the pSEEC for various separations in terms of their molecular weights (MWs) as well as p I s. For example, it was possible to separate the gel‐excluded proteins (BSA/IgG) as well as gel‐permeable and similar‐molecular‐weight proteins (Myo/Lys) by the pSEEC. Moreover, in the cases of Oval/Myo, which could be partially separated by size‐exclusion chromatography, the use of the pSEEC greatly improved the resolution and the separation became possible at high sample loading. The results indicate that the pSEEC technology is promising for preparative protein separations.