Surface modification and bioconjugation of FeCo magnetic nanoparticles with proteins

Magnetic Fe70Co30 nanoparticles with a cubic shape and a mean size of 15 ± 1.5 nm were fabricated using a magnetron–sputtering-based gas phase condensation deposition method. The particles had a high saturation magnetization of 220 emu/g, which is much higher than that of commercially available iron oxide nanoparticles. The FeCo nanoparticles were modified by 3-aminopropyltriethoxy silane and subsequently activated by glutaraldehyde, leading to successful attachment of aldehyde groups onto nanoparticle surfaces. Three proteins, namely streptavidin, PAPP-A antibody and Nectin-4 antibody, were immobilized on glutaraldehyde activated FeCo nanoparticles, and their loading levels were quantitatively evaluated. Our results show that loading capabilities are 95 μg of streptavidin, 128 μg of PAPP-A, and 125 μg of Nectin-4 antibody per milligram of FeCo nanoparticles, and that the three immobilized proteins retain their binding bioactivity. The protein–FeCo conjugates may find valuable applications involving magnetic separation and purification of proteins and cells, and the magnetic detection of biomolecules.