Methacryloylamidohistidine in Affinity Ligands for Immobilized Metal-ion Affinity Chromatography of Ferritin

Biotechnology and Bioprocess Engineering (Impact Factor: 1.11). 02/2011; 16(1):173-179. DOI: 10.1007/s12257-009-0162-4


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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