Methacryloylamidohistidine in affinity ligands for immobilized metal-ion affinity chromatography of ferritin

Biotechnology and Bioprocess Engineering (Impact Factor: 1.28). 01/2011; 16(1):173-179. DOI: 10.1007/s12257-009-0162-4

ABSTRACT 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.

Keywordsferritin–nanobiotechnology–nanosphere–IMAC–affinity chromatography

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    ABSTRACT: A novel metal-chelate affinity matrix utilizing N-acetyl cysteine as a metal chelating agent was synthesized. For this, magnetic Fe 3 O 4 core was coated with silver by chemical reduction. Then, these magnetic silver nanoparticles were covered with N-acetyl cysteine, and Fe 3? was chelated to this modified magnetic silver nanoparticle. These magnetic nanoparticles were characterized by SEM, AFM, EDX, and ESR analysis. Synthesized nanoparticles were spherical and average size is found to be 69 nm. Fe 3? chelated magnetic silver nanoparticles were used for the adsorption of ferritin from its aqueous solution. Optimum conditions for the ferritin adsorption exper-iments were performed at pH 6.0 phosphate buffer and 25 °C of medium temperature and the maximum ferritin adsorption capacity is found to be 89.57 mg/g nanoparticle. Ferritin adsorption onto magnetic silver nanoparticles was increased with increasing ferritin concentration while adsorption capacity was decreased with increasing ionic strength. Affinity of the magnetic silver nanoparticles to the ferritin molecule was shown with SPR analysis. It was also observed that the adsorption capacity of the magnetic silver nanoparti-cles was not significantly changed after the five adsorption/desorption cycles.
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