Purification of transferrin by magnetic nanoparticles and conjugation with cysteine capped gold nanoparticles for targeting diagnostic probes

Abstract

Transferrin is an iron binding glycoprotein actively involved in the growth and maintenance of cell cycle. The transferrin receptors expression is increased on growing cancer/tumor cells for absorption of iron through transferrin and participation in biological activity. In this study, a novel method for the purification of transferrin by using magnetic nanoparticles (MNP) is developed and compared with reported method. Magnetic nanoparticles were synthesized by co-precipitation method under hydrothermal conditions in the presence of ammonium hydroxide. MNP were characterized by FTIR, VSM, DLS, TEM, and SEM. Purified transferrin was characterized by SDS-PAGE, MALDI-TOF, ELISA, Western blot, and its activity was further confirmed by iron binding assay and receptor binding assays. Purified transferrin was also conjugated with cysteine capped gold nanoparticles (GNP) and characterized by UV-Vis spectra, TEM, DLS, and fluorescent spectrophotometry. Transferrin conjugated cysteine capped GNP used as a targeted fluorescent probe on gastric cancer, tumor tissue and MDA-MB 231 cancer cells to confirm transferrin receptor binding activity and application as diagnostic probe. The purified transferrin showed stability and activity up to 36 months. The results indicated that the synthesized superamagnetic MNP are good for the purification of transferrin. A good yield of transferrin was purified by this method, good quality and showed active biological activity. GNP conjugated transferrin has a potential to be used in cancer diagnosis as targeted diagnosis probe in vivo and in vitro. Experiments are underway for utilizing transferrin as carrier for targeting drug delivery.