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Preparation of Immobilized Metal Affinity Chromatographic Packings by Immobilization of Carboxymethylated Asparate (CM‐Asp) Based on Monodisperse Hydrophilic Non‐porous Beads and Their Application
Author(s) -
Gong Bolin,
Bo Chunmiao,
Wang Fuqiang
Publication year - 2010
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.201090203
Subject(s) - chemistry , dispersity , chromatography , chelation , metal , elution , metal ions in aqueous solution , zinc , copper , ligand (biochemistry) , affinity chromatography , nuclear chemistry , polymer chemistry , inorganic chemistry , organic chemistry , biochemistry , receptor , enzyme
The hydrophilic immobilized metal affinity chromatographic packing was prepared by immobilization of carboxymethylated asparate (CM‐Asp) as chelating ligand and Ni 2+ as center ion on the base of monodispersed, 3.0 µm non‐porous monodisperse poly(glycidylmethacrylate‐ co ‐ethylenedimethacrylate) (P GMA/EDMA ) particles. The retention behavior of proteins and the effect of pH on the retention in the range from 4.0 to 9.0 were investigated on both the naked and metal ion chelated columns. Four proteins were quickly separated in 2.0 min with linear gradient elution at a flow rate of 3.0 mL·min −1 by using the synthesized Ni 2+ ‐CM‐Asp‐P GMA/EDMA packings. The separation time was shorter than other immobilized metal affinity chromatography reported in the literature. The Ni 2+ ‐CM‐Asp‐P GMA/EDMA column was further investigated for the rapid separation and purification of copper‐zinc superoxide dismutase (Cu,Zn‐SOD) from the blood of pig in 3.0 min with only one step. The results obtained were satisfactory.