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KMID : 1094720110160010173
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Biotechnology and Bioprocess Engineering
2011 Volume.16 No. 1 p.173 ~ p.179
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Methacryloylamidohistidine in affinity ligands for immobilized metal-ion affinity chromatography of ferritin
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Uygun Deniz Aktas
Ozturk Nevra
Akgol Sinan
Denizli Adil
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Abstract
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A new metal-chelate adsorbent utilizing 2-methacryloylamidohistidine (MAH) was prepared as a metalchelating ligand. MAH was synthesized using methacryloly chloride and histidine. Monosize nanospheres with an average diameter of 450 nm were produced by emulsion polymerization of 2-hydroxyetylmethacrylate (HEMA) and MAH. Then, Fe3+ ions were chelated directly onto the monosize nanospheres. Mon-poly(HEMA-MAH) nanospheres were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and elemental analysis. Fe3+ chelated monosize nanospheres were used in ferritin adsorption from an aqueous solution. The maximum ferritin adsorption capacity of Fe3+-chelated mon-poly(HEMAMAH) nanospheres was 202 mg/g at pH 4.0 in acetate buffer. The non-specific ferritin adsorption on the monpoly( HEMA-MAH) nanospheres was 20 mg/g. The adsorption behavior of ferritin could be modeled using both Langmuir and Freundlich isotherms. The adsorption capacity decreased with increasing ionic strength of the binding buffer. High desorption ratios (> 95% of the adsorbed ferritin) were achieved with 1.0 M NaCl at pH 7.0. Ferritin could be repeatedly adsorbed and desorbed with the Fe3+-chelated mon-poly(HEMA-MAH) nanospheres without significant loss of adsorption capacity.
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KEYWORD
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ferritin, nanobiotechnology, nanosphere, IMAC, affinity chromatography
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