Immobilized metal affinity beads for ferritin adsorption

A new metal–chelate adsorbent utilizing N-methacryloyl-(L)-cysteine methyl ester (MAC) was prepared as a metal-chelating ligand. MAC was synthesized by using methacryloyl chloride and L-cysteine methyl ester dihydrochloride. Spherical beads with an average diameter of 150–200 μm were produced by suspension polymerization of 2-hydroxyethyl methacrylate (HEMA) and MAC carried out in an aqueous dispersion medium. Then, Fe3+ ions were chelated directly on the beads. Properties such as specific surface area, specific pore volume and ligand occupation were determined. The specific surface area of the beads was found to be 18.9 m2/g. The total pore volume was 2.8 ml/g and represented a porosity over 52%. The average pore size of the poly(HEMA-MAC) beads was 620 nm. Fe3+ -chelated beads were used in the adsorption of ferritin from aqueous solutions. Ferritin adsorption increased with increasing ferritin concentration. The maximum ferritin adsorption capacity of the Fe3+ -chelated poly(HEMA-MAC) beads (Fe3+ loading 0.81 mmol/g) was found to be 3.7 mg/g at pH 4.0 in acetate buffer. The non-specific ferritin adsorption on the poly(HEMA-MAC) beads were 0.4 mg/g. Adsorption behavior of ferritin could be modelled using both the Langmuir and Freundlich isotherms. Adsorption capacity decreased with increasing ionic strength of the binding buffer. Ferritin molecules could be adsorbed and desorbed five times with these adsorbents without noticeable loss in their ferritin adsorption capacity.

[1]  A. Denizli,et al.  A New Metal Chelate Affinity Adsorbent for Cytochrome c , 2008, Biotechnology progress.

[2]  M. Hynes,et al.  The kinetics and mechanisms of reactions of iron(III) with caffeic acid, chlorogenic acid, sinapic acid, ferulic acid and naringin. , 2004, Journal of inorganic biochemistry.

[3]  A. Denizli,et al.  Novel metal-chelate affinity adsorbent for purification of immunoglobulin-G from human plasma. , 2003, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[4]  S. Suen,et al.  Analysis of protein adsorption on regenerated cellulose-based immobilized copper ion affinity membranes. , 2003, Journal of chromatography. A.

[5]  E K M Ueda,et al.  Current and prospective applications of metal ion-protein binding. , 2003, Journal of chromatography. A.

[6]  M. Strumia,et al.  Influence of the morphology of poly(EGDMA-co-HEMA) on the chemical modifications and retention of O-phosphothreonine. , 2003, Journal of biochemical and biophysical methods.

[7]  A. Denizli,et al.  Heavy Metal Ion Adsorption Properties of Methacrylamidocysteine-Containing Porous Poly(Hydroxyethyl Methacrylate) Chelating Beads , 2002 .

[8]  J. Dybal,et al.  Purification of the specific immunoglobulin G1 by immobilized metal ion affinity chromatography using nickel complexes of chelating porous and nonporous polymeric sorbents based on poly(methacrylic esters). Effect of polymer structure. , 2002, Journal of chromatography. A.

[9]  G. Chaga,et al.  Twenty-five years of immobilized metal ion affinity chromatography: past, present and future. , 2001, Journal of biochemical and biophysical methods.

[10]  V. Gaberc-Porekar,et al.  Perspectives of immobilized-metal affinity chromatography. , 2001, Journal of biochemical and biophysical methods.

[11]  M. Y. Arica,et al.  Nonporous monosize polymeric sorbents : Dye and metal chelate affinity separation of lysozyme , 2000 .

[12]  M. Hearn,et al.  Characterisation by immobilised metal ion affinity chromatographic procedures of the binding behaviour of several synthetic peptides designed to have high affinity for Cu(II) ions. , 1999, Journal of chromatography. A.

[13]  M. Y. Arica,et al.  Affinity microspheres and their application to lysozyme adsorption: Cibacron Blue F3GA and Cu(II) with poly(HEMA‐EGDMA) , 1999 .

[14]  M. Y. Arica,et al.  Dye-ligand and metal chelate poly(2-hydroxyethylmethacrylate) membranes for affinity separation of proteins. , 1998, Journal of chromatography. A.

[15]  E. Roets,et al.  Analysis of kanamycin sulfate by liquid chromatography with pulsed electrochemical detection. , 1997, Journal of chromatography. A.

[16]  A. Denizli,et al.  Congo Red and Cu(II) carrying poly(ethylene glycol dimethacrylate-hydroxyethyl methacrylate) microbeads as specific sorbents Albumin adsorption/desorption , 1996 .

[17]  D. Nicholls,et al.  Metal binding and ferritin immunoreactivity in a high molecular weight fraction from rat brain. , 1996, Biochimica et biophysica acta.

[18]  G. Berthon.  Handbook of metal-ligand interactions in biological fluids, bioinorganic chemistry , 1995 .

[19]  G. Lindeberg,et al.  Importance of the alpha-amino group in the selective purification of synthetic histidine peptides by immobilised metal ion affinity chromatography. , 1995, Journal of chromatography. A.

[20]  T. Yip,et al.  Immobilized metal ion affinity chromatography , 1994, Methods in molecular biology.

[21]  U. Bahr,et al.  Immobilized metal affinity membrane adsorbers as stationary phases for metal interaction protein separation , 1994 .

[22]  J. Porath.  Immobilized metal ion affinity chromatography. , 1992, Protein expression and purification.

[23]  J. Porath,et al.  Surface topography of histidine residues: a facile probe by immobilized metal ion affinity chromatography. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[24]  M. N. Gupta,et al.  Immobilized Metal Affinity Chromatography without Chelating Ligands: Purification of Soybean Trypsin Inhibitor on Zinc Alginate Beads , 2002, Biotechnology progress.

[25]  A. Denizli,et al.  Diamine-plasma treated and Cu(II)-incorporated poly(hydroxyethylmethacrylate) microbeads for albumin adsorption. , 1999, Journal of biomaterials science. Polymer edition.

[26]  Elizabeth C. Theil.  Ferritin: structure, gene regulation, and cellular function in animals, plants, and microorganisms. , 1987, Annual review of biochemistry.