Preparation of magnetic metal-affinity sorbents and its use in a continuous flow system for cytochrome c adsorption
Abstract
A magnetic metal-chelate affinity adsorbent utilizing N-methacryloyl-(l_)-histidine methyl ester (MAH) as a metal-chelating ligand was prepared. MAH was synthesized using methacryloyl chloride and L-histidine methyl ester. Magnetic beads with an average diameter of 50-100 urn were produced by suspension polymerization of ethylene glycol dimethacrylate (EGDMA) and MAH carried out in a dispersion medium. Specific surface area of the magnetic beads was found to be 80 $m^ 2$/g. Elemental analysis of the magnetic beads for nitrogen was estimated as 70 umol MAH/g polymer. Magnetic beads were loaded with the $Cu^ {2+}$ ions directly via MAH for the adsorption of cytochrome c continuous flow system. The cytochrome c adsorption on the mag-poly(EGDMA-MAH) beads was 42 mg/g. $Cu^ {2+}$ complexing increased the cytochrome c adsorption significantly. The maximum cytochrome c adsorption capacity of the $Cu^ {2+}$-chelated beads (carrying 68 umol $Cu^ {2+}$ per gram of polymer) was found to be 197 mg/g at pH 8.0 in phosphate buffer. Cytochrome c adsorption decreased with increasing temperature. Cytochrome c molecules could be reversibly adsorbed and desorbed five times with the magnetic adsorbents without noticeable loss in their cytochrome c adsorption capacity. Cytochrome c adsorption decreased with increasing magnetic field. The binding isotherm was determined by scatchard initially followed by application of Hill equation to the data obtained, then binding constant and $n _H$ Hill coefficient were estimated. A magnetic metal-chelate affinity adsorbent utilizing N-methacryloyl-(l_)-histidine methyl ester (MAH) as a metal-chelating ligand was prepared. MAH was synthesized using methacryloyl chloride and L-histidine methyl ester. Magnetic beads with an average diameter of 50-100 urn were produced by suspension polymerization of ethylene glycol dimethacrylate (EGDMA) and MAH carried out in a dispersion medium. Specific surface area of the magnetic beads was found to be 80 $m^ 2$/g. Elemental analysis of the magnetic beads for nitrogen was estimated as 70 umol MAH/g polymer. Magnetic beads were loaded with the $Cu^ {2+}$ ions directly via MAH for the adsorption of cytochrome c continuous flow system. The cytochrome c adsorption on the mag-poly(EGDMA-MAH) beads was 42 mg/g. $Cu^ {2+}$ complexing increased the cytochrome c adsorption significantly. The maximum cytochrome c adsorption capacity of the $Cu^ {2+}$-chelated beads (carrying 68 umol $Cu^ {2+}$ per gram of polymer) was found to be 197 mg/g at pH 8.0 in phosphate buffer. Cytochrome c adsorption decreased with increasing temperature. Cytochrome c molecules could be reversibly adsorbed and desorbed five times with the magnetic adsorbents without noticeable loss in their cytochrome c adsorption capacity. Cytochrome c adsorption decreased with increasing magnetic field. The binding isotherm was determined by scatchard initially followed by application of Hill equation to the data obtained, then binding constant and $n _H$ Hill coefficient were estimated.
Source
Hacettepe Journal of Biology and ChemistryVolume
36Issue
3URI
http://www.trdizin.gov.tr/publication/paper/detail/TVRBME1EQXlNZz09https://hdl.handle.net/20.500.12418/1897
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