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Metal‐immobilized magnetic nanoparticles for cytochrome C purification from rat liver
Author(s) -
Perçin Işık,
Karakoç Veyis,
Ergün Bahar,
Denizli Adil
Publication year - 2015
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1347
Subject(s) - adsorption , nanoparticle , ionic strength , nuclear chemistry , chemistry , zeta potential , magnetic nanoparticles , emulsion polymerization , polymer , chromatography , polymerization , materials science , aqueous solution , organic chemistry , nanotechnology
Cu 2+ ‐immobilized magnetic poly(hydroxyethylmethacrylate‐ N ‐methacryloyl‐( l )‐histidinemethylester) (mPHEMAH) nanoparticles were prepared by surfactant‐free emulsion polymerization for cytochrome C (cyt C) purification from rat liver. Elemental analysis, atomic force microscopy, zeta sizer, and vibrating sample magnetometer were used to characterize mPHEMAH nanoparticles. In addition to these characterization steps, surface area, average particle size, and size distribution of mPHEMAH nanoparticles were determined. Quantity of immobilized Cu 2+ was measured using atomic absorption spectrophotometry. N ‐Methacryloyl‐( l )‐histidinemethylester and Cu 2+ content of mPHEMAH nanoparticles were 0.18 mmol/g polymer and 0.11 mmol/g polymer, respectively. Specific surface area of Cu 2+ ‐immobilized mPHEMAH nanoparticles was 1180 m 2 /g. Effect of initial cyt C concentration, pH, temperature, and ionic strength on cyt C adsorption onto Cu 2+ ‐immobilized mPHEMAH nanoparticles was investigated. Maximum cyt C adsorption capacity of Cu 2+ ‐immobilized mPHEMAH nanoparticles was 311.9 mg/g polymer. Maximum adsorption was obtained at pH 8.0 and 4 °C. Cu 2+ ‐immobilized mPHEMAH nanoparticles were used ten times with 4.1% decrease in adsorption capacity. In the last stage, Cu 2+ ‐immobilized mPHEMAH nanoparticles were used to purify cyt C from rat liver tissue, and the purity of desorbed fractions was controlled by SDS‐PAGE.