Chromatographic separation of proteins on metal immobilized iminodiacetic acid-bound molded monolithic rods of macroporous poly(glycidyl methacrylate-co-ethylene dimethacrylate).
Continuous rod of macroporous poly(glycidyl methacrylate-co-ethylene dimethacrylate) was prepared by a free radical polymerization within the confines of a stainless-steel column. The epoxide groups of the rod were modified by a reaction with iminodiacetic acid (IDA) that affords the active site to form metal IDA chelates used for immobilized metal affinity chromatography (IMAC). The efficiency of coupling of IDA to the epoxide-contained matrix was studied as a function of reaction time and temperature. High-performance separation of proteins, based on immobilized different metals on the column, were described. The influence of pH on the adsorption capacity of bovine serum albumin on the Cu2+-IDA continuous rod column was investigated in the range from 5.0 to 9.0. Purification of lysozyme from egg white and human serum albumin (HSA) on the commercially available HSA solution were performed on the naked IDA and Cu2+-IDA continuous rod columns, respectively; and the purity of the obtained fractions was detected by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry.
Luo
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Q
National Chromatographic R & A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian. zouhfa@mail.dlptt.ln.cn
Zou
H
H
Xiao
X
X
Guo
Z
Z
Kong
L
L
Mao
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X
D002846:Chromatography, Affinity / Q000295:instrumentation; D006863:Hydrogen-Ion Concentration; D007098:Imino Acids / Q000737:chemistry*; D008768:Methylmethacrylates / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D011506:Proteins / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D007098:Imino Acids; D008768:Methylmethacrylates; D011506:Proteins; C020347:GMA-EDMA; C008109:iminodiacetic acid