Magnetic nanoparticles (MNPs) have been used for purification of specific biomolecules form mixtures. The aim of this study is to develop a new, cheap, reusable, and magnetic-based material to purify the carbonic anhydrase (CA) enzyme in a short time with high efficiency. In the first part of this study, silica-coated iron oxide magnetic nanoparticles (Fe3O4@SiO2 MNPs) were obtained. Surface modification of Fe3O4@SiO2 MNPs was accomplished with 3-(4-Hydroxyphenyl) propionic acid (PA) and sulfanilamide (SA), respectively. SA is a selective inhibitor of CA, and it selectively binds to CA. The final particle was named Fe3O4@SiO2-PA-SA MNPs and characterized by SEM, TEM, XRD, and FT-IR. It was determined that the produced MNPs contained multicore, were smaller than 100 nm in size, and had a spherical morphology. The CA was purified from bovine blood hemolysate in a short time such as 2.5 h and in a simple manner. The maximum enzyme purifying capacity of MNPs was calculated as 13.87 ± 3.27 mg CA/g MNP. SDS-PAGE analysis was confirmed that high CA purification success was achieved.
Keywords: Carbonic anhydrase; Magnetic nanoparticles; Purification.
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