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Magnetic IDA‐modified hydrophilic methacrylate‐based polymer microspheres for IMAC protein separation
Author(s) -
Přikryl Petr,
Horák Daniel,
Tichá Marie,
Kučerová Zdenka
Publication year - 2006
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200600248
Subject(s) - iminodiacetic acid , methacrylate , glycidyl methacrylate , adsorption , chemistry , chromatography , elution , polymer , protein adsorption , microsphere , phosphate , magnetic nanoparticles , polymer chemistry , nuclear chemistry , materials science , chemical engineering , polymerization , inorganic chemistry , organic chemistry , chelation , nanotechnology , nanoparticle , engineering
Preparation of a new type of magnetic non‐porous poly(2‐hydroxyethyl methacrylate‐ co ‐glycidyl methacrylate) microspheres with hydrophilic properties containing coupled iminodiacetic acid (IDA) is described. The prepared microspheres were used for the immobilization of Ni(II) or Fe(III) ions to show their application in protein binding studies. Human IgG was bound to magnetic Ni(II)‐IDA‐modified microspheres and conditions of its adsorption and elution were optimized. Non‐specific binding of the protein to magnetic microspheres in the absence of Ni(II) ions was low. Fe(III) ions immobilized on magnetic IDA‐modified microspheres were used for the specific binding of porcine pepsin, as a model phosphoprotein. The ability of phosphate buffer to release the adsorbed enzyme from the microspheres and a low adsorption of the dephosphorylated protein indicate the participation of phosphate groups in the pepsin interaction. The elaborated method represents a rapid technique that can be used not only for the separation of proteins but also for analytical purposes.