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Electrophoretic affinity chromatography: method validation
Author(s) -
Liu Zheng,
Feng Shaohua,
Guo Suhui,
Shen Zhongyao,
Ding Fuxin,
Yuan Naiju
Publication year - 1998
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/(sici)1099-1352(199812)11:1/6<151::aid-jmr412>3.0.co;2-6
Subject(s) - chromatography , chemistry , elution , electrophoresis , adsorption , biomolecule , isoelectric focusing , matrix (chemical analysis) , desorption , gel permeation chromatography , analytical chemistry (journal) , biochemistry , organic chemistry , enzyme , polymer
Abstract A new method for preparative‐scale separation of biomolecules, electrophoretic affinity chromatography (EAC), is proposed in this paper. Separation by EAC is carried out in a long and ribbon‐like multicompartment electrolyser separated by membranes, in which the two central compartments are used for packing the gel matrix and for sample loading respectively. Next to the central compartments are the elution compartments and electrode compartments. The electric field is applied perpendicular to the fluid flow in the compartments. Adsorption and desorption steps may both be carried out in the presence of an electric field, which transports the target components into the gel compartment for adsorption and the impurities into the elution compartments for washing. After the adsorption step an elution solution is introduced and the product is released from the gel matrix and washed out. Separation of human serum albumin (HSA) from human serum gives HSA product of high purity, as demonstrated by isoelectric focusing analysis. The characteristics of electrophoretic binding of HSA on Blue Sepharose Fast Flow are examined. The preliminary results show that this new method has advantages in terms of high rate of mass transfer and ease of scaling up, which are of particular interest when large‐scale separation of biomolecules is considered. Copyright © 1998 John Wiley & Sons, Ltd.