Synthesis of orientedly bioconjugated core/shell Fe3O4@Au magnetic nanoparticles for cell separation

Orientedly bioconjugated core/shell Fe3O4@Au magnetic nanoparticles were synthesized for cell separation. The Fe3O4@Au magnetic nanoparticles were synthesized by reducing HAuCl4 on the surfaces of Fe3O4 nanoparticles, which were further characterized in detail by TEM, XRD and UV–vis spectra. Anti-CD3 monoclonal antibody was orientedly bioconjugated to the surface of Fe3O4@Au nanoparticles through affinity binding between the Fc portion of the antibody and protein A that covalently immobilized on the nanoparticles. The oriented immobilization method was performed to compare its efficiency for cell separation with the non-oriented one, in which the antibody was directly immobilized onto the carboxylated nanoparticle surface. Results showed that the orientedly bioconjugated Fe3O4@Au MNPs successfully pulled down CD3+ T cells from the whole splenocytes with high efficiency of up to 98.4%, showing a more effective cell-capture nanostructure than that obtained by non-oriented strategy. This developed strategy for the synthesis and oriented bioconjugation of Fe3O4@Au MNPs provides an efficient tool for cell separation, and may be further applied to various fields of bioanalytical chemistry for diagnosis, affinity extraction and biosensor.