A Novel Magnetic Adsorbent for Immunoglobulin‐G Purification in a Magnetically Stabilized Fluidized Bed

A novel magnetic poly(ethylene glycol dimethacrylate‐N‐methacryloly‐l‐histidinemethylester) [m‐poly(EGDMA‐(MAH)] support was prepared for purification of immunoglobulin G (IgG) in a magnetically stabilized fluidized bed by suspension polymerization. Elemental analysis of the magnetic beads for nitrogen was estimated as 70 μmol MAH/g polymer. Magnetic poly(EGDMA‐MAH) beads were used in the separation of immunoglobulin‐G (IgG) from aqueous solutions and/or human plasma in a magnetically stabilized fluidized bed system. IgG adsorption capacity of the beads decreased with an increase in the flow rate. The maximum IgG adsorption was observed at pH 6.0 for MES buffer. IgG adsorption onto the m‐poly(EGDMA) was negligible. Higher adsorption values (up to 262 mg/g) were obtained in which the m‐poly(EGDMA‐MAH) sorbents were used from aqueous solutions. Higher amounts of IgG were adsorbed from human plasma (up to 320 mg/g) with a purity of 87%. IgG molecules could be repeatedly adsorbed and desorbed with these sorbents without noticeable loss in their IgG adsorption capacity.

[1]  V. Muronetz,et al.  Isolation of antigens and antibodies by affinity chromatography. , 2003, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[2]  M. Vijayalakshmi,et al.  A potential set up based on histidine hollow fiber membranes for the extracorporeal removal of human antibodies , 2000 .

[3]  M. Radosevich,et al.  Affinity chromatography in the industrial purification of plasma proteins for therapeutic use. , 2001, Journal of biochemical and biophysical methods.

[4]  M. Y. Arica,et al.  Immobilization of glucoamylase onto spacer-arm attached magnetic poly(methylmethacrylate) microspheres : characterization and application to a continuous flow reactor , 2000 .

[5]  Z. Bílková,et al.  Oriented immobilization of galactose oxidase to bead and magnetic bead cellulose and poly(HEMA-co-EDMA) and magnetic poly(HEMA-co-EDMA) microspheres. , 2002, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[6]  M. Vijayalakshmi,et al.  Comparative affinity chromatographic studies using novel grafted polyamide and poly(vinyl alcohol) media , 1991 .

[7]  T. Nakane,et al.  Mannosylerythritol lipid, a yeast extracellular glycolipid, shows high binding affinity towards human immunoglobulin G , 2001, BMC biotechnology.

[8]  A. Denizli,et al.  Protein A-immobilized microporous polyhydroxyethylmethacrylate affinity membranes for selective sorption of human-immunoglobulin-G from human plasma , 2000, Journal of biomaterials science. Polymer edition.

[9]  P. Füglistaller,et al.  Comparison of immunoglobulin binding capacities and ligand leakage using eight different protein A affinity chromatography matrices. , 1989, Journal of immunological methods.

[10]  Y. Sun,et al.  Protein adsorption equilibria and kinetics to a poly(vinyl alcohol)-based magnetic affinity support. , 2001, Journal of chromatography. A.

[11]  M. Vijayalakshmi.  Pseudobiospecific ligand affinity chromatography , 1989 .

[12]  Yan Sun,et al.  Application of Magnetic Agarose Support in Liquid Magnetically Stabilized Fluidized Bed for Protein Adsorption , 2003, Biotechnology progress.

[13]  M. Burns,et al.  Overcoming support limitations in magnetically stabilized fluidized bed separators , 1991 .

[14]  P. Blond,et al.  Purification of immunoglobulins G by protein A/G affinity membrane chromatography. , 1999, Journal of chromatography. B, Biomedical sciences and applications.

[15]  K. Kroner,et al.  Surface-modified membranes as a matrix for protein purification. , 1992, Journal of chromatography.

[16]  A. Denizli,et al.  Polyhydroxyethylmethacrylate-based magnetic DNA-affinity beads for anti-DNA antibody removal from systemic lupus erythematosus patient plasma. , 2001, Journal of Chromatography B: Biomedical Sciences and Applications.

[17]  G. Fassina,et al.  IRIS 97: an innovative protein A-peptidomimetic solid phase medium for antibody purification. , 1998, Journal of molecular recognition : JMR.

[18]  I. Ishigaki,et al.  Adsorption and elution of bovine gamma-globulin using an affinity membrane containing hydrophobic amino acids as ligands. , 1991, Journal of chromatography.

[19]  Peter Dunnill,et al.  Non-porous magnetic chelator supports for protein recovery by immobilised metal affinity adsorption , 1996 .

[20]  Z. Su,et al.  Protein A immunoaffinity hollow fiber membranes for immunoglobulin G purification: experimental characterization. , 1995, Biotechnology and bioengineering.

[21]  Cristina Martín,et al.  Synthesis of a Novel Magnetic Resin and the Study of Equilibrium in Cation Exchange with Amino Acids , 2004 .

[22]  C. Fee,et al.  Preparation of magnetically susceptible polyacrylamide/magnetite beads for use in magnetically stabilized fluidized bed chromatography. , 1997, Biotechnology and bioengineering.

[23]  R. Carbonell,et al.  Affinity chromatographic screening of soluble combinatorial peptide libraries. , 1999, Biotechnology and bioengineering.

[24]  Y. Sun,et al.  A Novel Magnetic Affinity Support for Protein Adsorption and Purification , 2001, Biotechnology progress.

[25]  M. Strumia,et al.  Preparation of adsorbents applicable to pseudobiospecific ligand affinity chromatography using different spacers and ligands , 1997 .

[26]  M. Burns,et al.  Dried calcium alginate/magnetite spheres: A new support for chromatographic separations and enzyme immobilization , 1985, Biotechnology and bioengineering.

[27]  M. Vijayalakshmi,et al.  Experimental kinetic aspects of hollow fiber membrane-based pseudobioaffinity filtration: process for IgG separation from human plasma , 1996 .