Cytochrome c adsorption in a continuous flow system by using Cu(II)-chelated magnetic affinity particles

Abstract

Background and methodology: In the current study magnetic poly(ethylene glycol dimethacrylate-N-methacryloyl-(L)-histidine methyl ester) poly(Egdma-Mah)) was used as a metal chelated affinity particles. Cu2+ ions loaded directly to MAH ligand of magnetic particles for the adsorption of cytochrome c (Cyt c) in a continuous flow system. Results: The maximum Cyt c adsorption capacity on the magnetic particles and Cu2+-chelated magnetic particles were 42 mg/g and 197 mg/g in phosphate buffer (pH 8.0), respectively. Cu2+ loading increased the Cyt c adsorption capacity, significantly. Cyt c adsorption capacity decreased with increased temperature and with increasing magnetic field. According to reusability studies Cyt c molecules could be reversibly adsorbed and desorbed five times. The binding isotherm was determined by scatchard analysis followed by application of Hill equation to the data obtained, then binding constant and nH Hill coefficient were calculated.