This investigation indicated an efficient procedure to purify human carbonic anhydrase II (hCA II) enzyme through sulfanilamide-functionalized (γ-Fe2O3-CPTES-SA) magnetic nanoparticles (MNPs), where synthesis of Fe3O4 MNPs was carried out using co-precipitation reaction. Next, 3-chloropropyltriethoxysilane (CPTES) was used to modify Fe3O4 nanoparticles and lastly, the surface of the nanoparticles was functionalized with sulfanilamide (SA) as a carbonic anhydrase ligand/inhibitor for binding to hCA II. The characterization of the synthesized nanoparticles was performed using various techniques. These characterization methods revealed that the MNPs were effectively coated with CPTES and SA, and the average diameter of the nanoparticles was approximately 21 nm. The possibility of interaction of γ-Fe2O3-CPTES-SA nanoparticles with hCA II was studied via multi-spectroscopic techniques. The protein isolated by this single-step procedure had high purity as confirmed by a single band on SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) which was also active after purification. By focusing on their drug loading capacity, and increasing their specificity and affinity to target CA-expressing cancer cells, the synthesized MNPs may dramatically impact the treatment of cancer and become suitable for clinical use in the near future.Communicated by Ramaswamy H. Sarma.
Keywords: Magnetic nanoparticles; human carbonic anhydrase II; protein purification; sulfanilamide.