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Superior magnetic monodisperse particles for direct purification of immunoglobulin G under magnetic field
Authors:Nilay Bereli
Institution:Department of Chemistry, Biochemistry Division, Hacettepe University, Ankara, Turkey
Abstract:Monodisperse magnetic acrylate based particles (5.0 µm in diameter) containing histidine were synthesized using a modified suspension polymerization method for the purification of immunoglobulin G from human plasma in a magnetically stabilized fluidized bed. N-methacryloyl-(L)-histidine methyl ester (MAH) was used as pseudo-specific ligand/co-monomer. MAH content of the magnetic particles was calculated as 55.3 µmol MAH/g polymer using elemental analysis. Immunoglobulin G binding amount of the magnetic particles decreased with increase of the flow-rate. The maximum immunoglobulin G binding was observed at pH 7.4 (phosphate buffer). Immunoglobulin G binding amount onto the magnetic poly(ethylene glycol dimethacrylate) mPEGDMA] particles was found to be almost negligible due to the hydrophilic polymer structure. High binding values were obtained from aqueous solutions (1646 mg/g). Higher immunoglobulin G binding was observed when human plasma was used (2169 mg/g). Purity of the separated immunoglobulin G from human plasma was found to be 87%. Magnetic PEGDMAH particles could be used many times without significant loss in protein binding amount.
Keywords:Affinity chromatography  magnetic PEGDMA particles  affinity separation  antibody purification
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