- 期刊
A Poly (2-hydroxyethyl Methacrylate)-based Immobilized Metal Affinity Chromatography Adsorbent for Protein Purification
並列摘要
Poly (HEMA) microbeads were prepared by suspension polymerization of 2-hydorxyethylmethaerylate and ethyleneglycoldimethacrylate (EGDMA). The water content, ligand density, and selectivity for poly (His)-tagged D-hydantoinase of the poly (HEMA)-based adsorbents were affected by tile concentration of EDGMA used during polymerization. The Ni(Ⅱ)-loaded poly (HEMA) adsorbent exhibited an adsorption capacity of 1.0 mg/g for poly (His)-tagged D-hydantoinase under optimal conditions with buffer containing 100-300 mM NaCl at pH 6.0. One-step purification protocol with the adsorbent gave a purity of at least 92%. The adsorption capacity of adsorbent declined by 54% after 7 cycles, due to the leaching of Ni(Ⅱ) from the adsorbent. However; upon regeneration the adsorption capacity can be restored. Given the ease of preparation and the chemical and microbial resistance, the poly (HEMA)-based IMAC adsorbent could be a promising substitute for the polysaccharide-based IMAC adsorbents.
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