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Preparation of Immobilized Metal Affinity Chromatographic Packings Based on Monodisperse Hydrophilic Non‐porous Beads and Their Application
Author(s) -
BO ChunMiao,
GONG BoLin,
HU WenZhi
Publication year - 2008
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.200890163
Subject(s) - chemistry , chromatography , iminodiacetic acid , chelation , elution , dispersity , trypsin , metal ions in aqueous solution , metal , affinity chromatography , glycidyl methacrylate , polymer chemistry , inorganic chemistry , organic chemistry , polymerization , enzyme , polymer
Three hydrophilic immobilized metal affinity chromatographic packings for HPLC have been synthesized by chemical modification of 3.0 µm monodisperse non‐porous poly(glycidyl methacrylate‐ co ‐ethylenedimethacrylate) (P GMA/EDMA ) beads. The retention behavior of proteins on the metal ion chelated columns loaded with copper(II), nickel(II) and zin(II) ion was studied. The effect of pH on the protein retention was investigated on both the naked and metal ion chelated columns in the range from 4.0 to 9.0. Four proteins were quickly separated in 3.0 min with linear gradient elution at a flow rate of 3.0 mL/min by using the synthesized Ni 2+ ‐IDA (iminodiacetic acid) packings. The separation time was shorter than other immobilized metal affinity chromatography reported in the literature. Purification of lysozyme from egg white and trypsin on the commercially available trypsin was performed on the naked‐IDA and Cu 2+ ‐IDA columns, respectively. The purities of the purified trypsin and lysozyme were more than 92% and 95%, respectively.