Multifunctional polymer-derivatized γ-Fe2O3 nanocrystals as a methodology for the biomagnetic separation of recombinant His-tagged proteins

• Published in: Journal of magnetism and magnetic materials 2008-10, Vol.320 (19), p.2339-2344
• Main Authors: SHUKOOR, M. I, NATALIO, F, TREMEL, W, TAHIR, M. N, DIVEKAR, M, METZ, N, THERESE, H. A, THEATO, P, KSENOFONTOV, V, SCHRÖDER, H. C, MÜLLE, W. E. G
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Condensed matter: electronic structure, electrical, magnetic, and optical properties; Diamagnetism, paramagnetism and superparamagnetism; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Magnetic properties and materials; Physics; Proteins
• ISSN: 0304-8853
• DOI: 10.1016/j.jmmm.2008.04.160

Multifunctional polymer-derivatized superparamagnetic iron oxide (gamma-Fe2O3) nanoparticles were prepared for biomagnetic separation of histidine-tagged recombinant proteins building up a faster and efficient method for protein separation by making use of their intrinsic magnetic properties. Using polymer bound gamma-Fe2O3 nanocrystals, a 6X histidine-tagged recombinant protein (silicatein) with a molecular weight of 24 kDa has been isolated and purified. The supermagnetic iron oxide nanocrystals were characterized by transmission electron microscopy (TEM), high-resolution TEM (HRTEM), SQUID and Mossbauer and the polymer functionalization of the gamma-Fe2O3 nanocrystals was monitored by UV-vis spectroscopy and light microscopy. Protein immobilization and separation was monitored using immunostaining techniques and gel electrophoresis, respectively.